STUDIES ON THE TAXONOMY AND BIOLOGY OF SOME DIPLOGASTRID SPECIES FROM INDIA

THESIS SUBMITTED FOR THE AWARD OF THE DEGREE OF TDoctor of Pbili)!gopbp IN ZOOLOGY

BY GAZALA YOUSUF

DEPARTMENT OF ZOOLOGY ALIGARH MUSLIM UNIVERSITY ALIGARH -202002, INDIA .- C1S 2013 .c~5 iad f rh, BOO

13 ,6V 2014

IIIIIIII III IIIII illill IIIII lillil!l T9005 , IRFAN AHMAD Ph.D. DEPARTMENT OF ZOOLOGY Professor Aligarh Muslim University, Aligarh — 202002, INDIA ahmadirfi c,gmaiI.com

Date: ......

Certificate

This is to certify that the research work embodied in the thesis entitled, "Studies on the Taxonomy and Biology of some Diplogastrid species from India" by Ms. Gazala Yousuf is original and was carried out under my supervision. I have permitted Ms. Gazala to submit it to the Aligarh Muslim University, Aligarh in fulfillment of the requirements for the degree of Doctor of Philosophy in Zoology.

IRFAN ARMAD (Supervisor) )lcknow(ecfgements

6vu to deep re(ereucc to the Altrtig(ty A(Kih, the most 6eneficia(and mercifu( ,whose benign blessings led me to complete this wcvorl,and produce it in its present fonn.

From the core of my heart, I express my deep sense of gratitude to my supertasor Prof Irfan Ahmad, Chairman, •Department of ."oolojy, aI' -Dean, 'Facu(ty of Life Sciences, /1(efjarh :tfus(unt Z'nicersity, A(igarfr for his eter-fasting important ad.ices, valuable suggestions, stimulating discussions, constructice criticis►n, sense of perfection and precision tchic( enabled me to complete this tror1,. Inspite of his tight departmental commitments he (dept his door wide open for ire. I(is affectionate instinct and constant encouragement were always a boon to me. It has been an honour to to orry with him.

I also tharva ,Prt!..11. S(antun ,jairajpuri r1'\;5 ,Senior, Scientist I, Prof •Rasi►n )tlztmad Prof Qudsia lahsee►t and Pro,;%%faha(aga (Youdhary for their valuable advices and kind suggestions.

Helping hands extended by my seniors and (a6 colleagues, Dr. Moorus ,Sa6a(e, Or Md ufahamood, ,Dr. Gauraz Singh, ;Mr. (Puneet Kumar e Wrs..A'adia Sufyart are highly acknoufedged here with thanIs.

I am also t(tanGfu(to my seniors Dr. ifd Baniyamuddirt, Dr. S(ikha Al fa-wat, Dr. 'Vyay Yikram Singh, Sots. Z'zma Tauheed, wr. i4d )lsif, Efs. Yaf&a 'Wustagirn, .Ms. Sumaya)4had and 11s. Zarrn Imran for their pleasant company, help and support.

Special thanks are due to 'Late) _Mrs. '1"abinda Ivusrat for her moral support, suggestions and above all good company. She a as more than just a friend and the coif she (eft in my heart can serer be fiffed:

I tuou(d file to proride a token off thanks to my friends Azf a, Seema, ifuma, Sadaf, Xqzia, %al wish and'7' a.ia, a (to though not a part of this research i ork. but supported me to an a ytrerne extent which I can't forget.

?flanks are also due to my class-mates ;Ifafino, 7alak, 'Farha, 'Yusra and Waseem for their help and support.

My words of thanks will remain incomplete if I forget to mention the names of my friends 'Fatima, 7fumaira andSumaira, who were oath me during the ups and downs of my research, gave me their precious time, patient hearing and kindness during the days I really found difficult to carry out my workand sorted out my pro6iems.

Last but not the (east, I wouldG&e to thank my Parents, my elder Brothers Mr. 'Khal dlousuf a ;Nr. ;NubashIr ;Yousu f my Sister-in-law ;%1rs.:Naroof Aaiir for their unconditional support, loco and never ending care. I am equally thankfu( to my little niece Tahura BtnteN,ha(id forgiBang me cheers of my life.

Gaala Yousuf CONTENTS

Page

Introduction I

IIistorical Background 7

Taxonomic revic« of free-living nematodes 7

Re% ie%% on biology:) of free-living nematodes 30

Materials and Methods 42

Systematics 46

Order Rhanditida 46

Suborder 1)iplogastrina 46

Superfamii> Diplogastroidea 47

amih Diplogasteroididae 48

SuhfamiI 1)iplogasteroidinae -48

Genus (;offurtiu 48

Go fiirtiu bcrlchus sp.n. 49

Subfamily Rhabditolaiminae 55

Genus Rhandliticloicic.s

Rhuhditidoi(l s hapillutu.t 5 5

Family Diplogastridae 61

Subfamily Diplogastrinae 61

Genus Aerosviehus 6I

Acrostichuc nudicupitutus 62

Genus Butlerius 68

Butlerius micuns 68

Genus Iiplogu.vtrellus 74 I Diplogasirellus curvatus sp. n. 74

Diplogastrellus neograciloides sp. n. 80

lli1,lr,ga.c•trdlIus ka.shmerieasis sp. n. 86

Diploc astrellus loktakai sp. n. 92

Diploga.strellus gracilis 98

Genus I'serrckuvusticlt,c.s gen. n. 103

I'.%rtnk,c'ruslirhuo ,uu.sal sp. n. I04

Subfamily Ihmaniellinac 111

Genus Dernaniellu 1 l 1

Demaniella keirakensis l 1 1

Family Neodiplogastridae 117

Subfamily Neodiplogastrinae 117

Genus Fictor 118

bictor suptilts sp. n. 118

Fictor 1aj2giruac!u sp. n. 125

Genus Koerneria 131

Koerl1e!ia/llicaudala 131

Genus 1!fononc1roirlc.c 137

Afononchoick's (enuispu•u!a sp. n. 137

.%fononchoide.c latiguhernacula sp. n. 144

Afononc/,oicic'.s .sn•uitu.c 1411

Genus ()igolaimella 156

Oigolaimella breviguhernacula sp. n. 156

Oigolaiacl!a longic•auda 162

Genus I'ris1ion{lu.c 167

l'risiionc/ics stercnrari.s sp. n. 167

I mil%"I\IL~pharyngidae 174

Genus 7}ioplttol•itr 17.1

71 lop/tarr?rr foctidu 17.1

SuperfamiIy Cylindrocorporoidea 180

Family C'\ lindrocorporidae Goodey 180

Genus itivctoluiiinc.s I80

:I c lo/arinuLc 1a1illalrt.s sp. n. 18

:11i ctolainucs oreodoxus sp. n. 187

Biology of Alttlolai,nus oreodoxity sp.n. 193

F.mhryogcncsis 193

Post-embryogenesis 195

Discussion 204

Summar\ 209

1(eI renccs 214 / ice..` l The nematodes are highly diversified lower invertebrates. The name nematode comes from the Greek word 'nematos' which means 'thread' and 'eidos' meaning

'form'. Nematodes are putatively the most species-rich animal phylum. The have various life styles and occur in a variety of habitats, ranging from free-li\ ing in marine, fresh-waters or in soil or parasites of different kinds of plants and animals.

I he\ are found in almost all t\ pes of habitats and occur in unimaginable numbers, sires, shapes and structures. Nematoda is a \er\ successful group of animals, four of even five multicellular animals on the planet are nematodes (Gunapala et a/., 1998).

They are present almost everywhere such as crop fields, in sand dunes, sediments beneath the ocean floor. freshwaters, in plants, animals and even in humans. This croup is perhaps the most abundant group among multicellular animals on earth. They are di\erse metazoans with an estimated total number of a million species

(l.ambshead. 2004). A single acre of soil from the arable lands may contain more than three billion nematodes. while the marine beach sand ma\ contain more than one and a half billion nematodes per acre.

Their abundance reaches many million individuals per m- in soil and bottom

sediments of aquatic habitats and a handful of soil may contain more than 50 different

species of nematodes. Sur e\'s of benthic sediments suggest that the total species

number for marine nematodes may exceed one million (1.ambshead. 1993:

Lambshead, 2001) with only a f'c~w thousands described in the scientific literature

(Malakhov, 1994: De Lc \ & Blaxter. 2002). In terrestrial systems, nematode diversity

appears to be under-reported (Lawton et al., 1998), 'ith for example. only about 200

species of soil nematodes being described from the British Isles (Boag & Yeates.

1998). The maximum number of nematode taxa described from a single soil site is

228 from a prairie in Kansas. USA (Orr & Dickerson. 1966: Boar & Yeatcs, 1998).

Their size too is extremely variable ranging from less than I00 fun (GrceJicllu minutu?n) to greater than 8 metres (Pla(enton'!na gigantissimn) discovered in the placenta of a sperm hale. Several Lon i(lHrr►s species exceed 10mm in length.

From an environmental point of view. nematodes are part of nearly all ecos'stems in their roles as bacterivores, herbivvores, parasites of animals and plants, and consumers of dissolved as well as particulate organic matter. They have critical roles in flow of energy and cycling of' nutrients. From an anthropocentric point of view, they parasitize humans and plants. domestic and \\ild animals and they can serve as indicators of environmental change. With regard to health, for example. a large proportion (22%) of the human population is parasitized by the intestinal

parasite Ascaris• lumbricoules (Crompton, 1988). Considering their impact on crops.

McCarter (2009) estimated a global total loss of $118 billion for 2001, of which

nearly halt' was related to only two crops. rice and maize. This being so, it is

remarkable that they are among the least studied, with only close to 26.000 (< 3% of

estimated) species described to date (I lugot etal.. 2001: 1-lallan. 2007).

The deepest oceans, freezing polar seas, hot water springs, arid deserts, alpine

lakes, mountain tops and the frozen Antarctica, represent the hostile climates of

«orid. but surprisingly nematodes are not only present but quite in abundance in all

these ecosystems. Tropical rain-forests and also the subtropical areas of the world

have tremendous nematode diversity. Soil nematode diversity has been one of the

main foci in ecological researches. Nematodes occupy a central position in the soil

food %%eb and play significant roles in ecological processes such as nitrogen cycling

and plant growth patterns (Neher, 2001). Their abundance, trophic classification and

various indices such as Shannon-Weaver index, dominance index, enrichment index,

structure index, channel index all have been used to varying degrees to assess soil

management (Berkelmans et al.. 2003: Bongers. 1990: Ferris et al., 2001, Yeates,

1999). Soil nematodes offer great potential for use as indicators of biodiversity and

2 ecological stability, and for assessing the impact of changing land use on soil conditions. Various studies have shown that difacrent nematodes have different life spans. different reproductive and survival capacilN. different positions in the food

web, respond difterentl' to environmental changes and can be used as ecological

bioindicators (Freckman. 1982: Bongers. 1990) and hio-control agents (Jairajpuri &

Hi l cram i. 1990: Ahmad. 1990).

Nematodes are ubiquitous and are diverse in feeding behaviour and life

strategies. The diversity of feeding behaviour and some intrinsic properties result in

the complex biotic interaction between soil nematodes and other soil biota which

significantl\ affects the ecosystem functions. Nematodes are the integral part of the

food web in the sub-terranean ecosystem and are divided into different trophic groups

such as plant feeders, hyphal feeders, bacterial feeders. substrate ingesters. animal

predators. unicellular eukaryote feeders. dispersal or infective stages of animal

parasites and omni\ores based on their diverse feeding habits. These groups comprise

different trophic specializations and are sensitive to soil disturbances and nutrient

enrichment. The ecological classification of terrestrial nematodes has usuall\ been

based on feeding biology: trophic function and on the life strategies: colonizers vs

persisters (Bongers. 1990). The nematode ecologists generally recognize following

major trophic groups among soil inhabiting forms on the basis of their feeding

apparatus:- i) herbivores, ii) fungivores. iii) bacteriovores. iv) predators and v)

omnivores (Yeatcs ei al.. 1993: Yeates, 1998: Yeates & Bongers, 1999).

Nematodes feeding on vascular plants using a stomato sty let, odontostyle or

onchiosty Ie are called as herbivores. Fungivores puncture the fungal hyphae by a

stylet with a narrow lumen. t3acteriovores feed on any prokaryotic food source

through a narrow or broad stoma. Predators feed on other nematodes or small

invertebrates such as protista. rotifers and enchvtraeids. They may he either ingesters

3 or piercers. Omnivores feed on a range of food sources. It is usual to restrict the term omnivore to some dorvlaims.

l3acteriovorous nematodes are ke\ microtaunal grazers that regulate ecological processes of decomposition and nutrient cycling thereby indirectly of ecting primary

production. An increase in the abundance of' this group often enhances microbial activit\ in soil with a non-limiting supply of' nitrogen (\\'asile\wska. 1998).

Bactcrivorous nematodes are better indicators of the rate of decomposition of organic

matter than the abundance of bacteria because. through their higher position in the

food chain, they integrate both biotic and abiotic factors (Griffiths et al.. 1994:

Wasilewska. 1998). Other evidences indicate that bacteriovorous nematodes can

increase soil mineral nitrogen availability, crop growth and nitrogen uptake in

microcosm and field experiments (Ferris et al.. 1997; Ingham et a/.. 1985). Thus any

change in land management that affects bacteriovorous soil nematodes has the

potential to influence critical ecological processes (Yeates & King. 1997).

Nematology gained attention and recognition only in 20`x' century, but our

knowledge of a few species of nematodes of medical importance dates back to

Paps rus Ehers (Circa 15,00 BC). The intestinal round worm (Ascaris lun hricoide.$).

tilarid (H'uchereria hancrofti) and Guinea worm or tier\ serpent of Moses

(Dracunculus medinensis), an 80 cm long nematode that prefers to live under human

skin were already known to the ancient man. Avicenna (('irca 1000 AD) prescribed a

treatment for the pin orms (Enierohius verrnicularis) which is used even today.

Knowledge of free-living nematodes dates back to 1656 when l3orellus for the

first time observed Turbatrix aceti the 'vinegar eels'. Animal parasites received much

attention by the ancient scientists than plant parasitic nematodes which were less

conspicuous at that time. Needham (1743) solved the "riddle ol' cockle" when he

crushed one of the diseased wheat grains and observed "Aquatic Animals" or serpent-

4 like worms, the first plant parasitic nematode (Anguina tritici) which were later named Fihrio tritici (Steinbuch. 1799). Muller (1783) described several species of free-living freshwater nematodes. Nematode taxonomy Further developed and a remarkable progress \'. as observed in the middle of 19`x' center\ . Research on the tree

Iii mu nematodes de eloped rapidl\ after 1980. when the possibilities of using nematodes in environmental studies became apparent.

Dujardin (I 845) first established the genus Rhabcliiis with R. terriculcr as its tNpe species. However. it was not clearly defined until more than one hundred \-cars later (Dougherty, 1955). Orley (1880) tried to fit the genus Rhabditis into the system of Nernatoda and proposed a family Rhabditidae for the genera Anguillula,

C'ephalobus. O.rvuri.c. Rhabditis and Teratocephalu.c and placed this family in the

higher category 'Rhabditiformae' which formed a connecting link between free-living

and animal-parasitic nematodes. lie provided information about biology and

taxonomy of Rhahc/iti.c and also provided a key ti)r the species identification. 1)e Man

(1876, 1880. 1884) added many new species and accepted thirty-seven species in

Rhahclit ic.

Around 1850. marine biologists began to recognize nematodes: they started

studies on the nematodes of Iceland (Leuckart, 1849). the Mediterranean (E;berth,

1863), the English coast (Bastian, 1865), the coast of Brittany (Villot, 1875) and on

nematodes collected by various expeditions (Von I.instow, 1876). Freshwater

nematodes received further interest around 1890 with the papers of Daday (1 897) on

the Hungarian fauna. Dujardin (1845). Bastian (1865), Schneider (1866), de Man

(1884). Daday (1905) and Maupas (1900) were the pioneers of the field. Early work

on the free-living nematodes included careful descriptions of Enohlus. (hncholaimu.s,

Rhabditis and Dorvlaimus• (E)ujardin. 1845). 1Ic was first to recognize the close

relationship of free-living and plant parasitic nematodes. Bastian (1865) also made a

5 significant contribution in the field of Nematology. lie catagorized the free-living nematodes into soil, fresh «ater and marine forms and described 100 new species of

30 genera in which 23 were new to science. in a single paper. De Man (1876-1927)

listed eight families of free-living nematodes. De Man's (1884) formula for denoting

morphometric measurements of nematodes is universally used in taxonomy till date.

Cobb. a contemporary of de Man. is considered. the Father of Nematology. Ile placed

nematodes under separate Phylum Nemata. Significant changes in classification were

proposed b~' Cobb (1920), De ('oninck (1965). Maggenti (1963. 70). Siddiqi (2000),

Sudhaus (2003), De Ley (2002, 2003) and by Andrassy (1976, 84. 2005). Cobb

(1913) proposed the genus Diploscapter for Rhabcliiis corunata. Micoletzky (1922),

in his monograph on the non-marine free-living nematodes, listed 142 valid genera

and 931 species. Chitwood (1933) established the Order Rhabditida for

bacteriophagus rhabditids. In Chitwood's (1933, 37) classification. 'Nematoda' was

treated as a ph\ lum vv ith two classes, 'Phasmidia' and 'Aphasmidia', based on

presence or absence of phasmids. The terms Secernentea and Adenophorea \\ere

introduced by Chit\~ood (1958) \vho proposed the system of classification of

nematodes including free-living and parasitic nematodes.

0

Taxonomic review of free-living nematodes

The classification of Rhabditida went through much alterations and have been modified several times b man\ taxonomists. Chitwood and Chitwood (1950) assigned four suborders including face-living and parasitic forms under Rhanditida:

Rhabditina. "I vlenchina. Strongylina and .Ascaridina. Goodey (1963) distinguished three superfamilics under the suborder Rhabditina: Diplogastroidea,

Pseudodiplogastroidea and Rhabditoidea. fie (I.c.) separated cephalobids,

panagerolairnids and bunonematids under family levels. Andrassy (1974) was the first

who separated Teratocephalina in a suborder rank and added Cephalobina, Rhabditina and Diplogastrina as further suborders. lie also listed t~'o. six, six and live families

within these suborders respectively. Lorenzen (1981) considered Rhabditida to

comprise of the two superfamilies: Rhabditoidea and Diplogastroidea. He placed five

and two families under Rhabditoidea and Diplogastroidea respectively. In the same

year. Maggenti (1981) grouped the rhabditid nematodes under two suhorders.

Rhabditina and Cephalobina and assigned three and five superfamilies, respectivel`.

Malakho\. Rvzhiko\ and Bonin 1982) proposed three suborders: Cephalohina.

Rhabditina and 1)rilonematina. In 1983. Inglis distinguished four suborders under

Rhabditida: Rhabditina, Angiostomatina, Bunonematina and Myolaimina. De Ley and

Biaxter (2003) divided the free-living Rhabditida into three suborders: Myolaimina.

Tylenchina and Rhabditina. Under Rhabditina they placed three infraorders:

Bunonematomorpha. Diplagastromorpha and Rhabditomorpha which include two.

eight and four families, respectively.

Recently. Andrassv (2005) split up the Rhabditida into five suborders:

Teratocephalina Andrass~, 1974, Cephalobina Andrassy. 1974, Myolaimina Inglis,

1983. Rhabditina Chitwood. 1933 and Diplogastrina hvlicoletzky. 1922. lie assigned

one family to the suborder l eratocephalina which includes t~'o genera. Under the

7 suborder Cephalobina he assigned three superfamilies x%hich include seven families, eighteen subfamilies and sixt\-one =enera. Mvolaimina includes only one family having a single genus with nine species. He divided suborder Rhabditina into two supertamilies within which he assi_ned six families and accepted Ii t\ genera and 329 species. He divided Diplogastrina into three superfamilies and nine families.

Andrassy contributed cxtensi\cly to the taxonomy of major groups of terrestrial and freshwater nematodes. Ile described more than 500 taxa of nematodes and almost 39 taxa were named after him. Besides his voluminous contributions to nematode taxonom\ and systematics, he has an enormous influence on soil and nematode ecology. He published keys for identification. proposed and raised higher taxa. amended and put forth classification schemes besides authoring valuable books including the extremely useful compilation. 'Klccsse Nemcnoda (1984) based on the diagnosis of orders of :\ raeolaimida. Enoplida. Chromadorida. Monhysterida, and

Rhabditida and their subordinate taxa. This hook exercised major influence on the direction of nematode ecology in that it bridged the gap between nematode taxonomy

and soil ecology. lie also authored "the nematode fauna of Hungary" in three

volumes. The first volume (2005) deals with the orders Monhvsterida, Areolamida.

Chrornodorida and Rhabditida. The second volume (2007) deals with the orders

Aphelenchida. Tvlenchida and Enoplida. The third volume (2009) deals \\ ith the

orders Triplonchida. Mononchida and Dorylaimida.

The Diplogastrids are a heterogeneous group of nematodes. Tlie are

predominantly terrestrial found in dung. compost, leaf litter, slime flux, manure,

decomposing plants or any other organic rich environment. Some species live in

association with insects, some inside insects, others live in the tunnels of, hark beetles

and a few are aquatic. [)iplogastrid nematodes are considered to be a

monophyletically developed group (Sudhaus, 2003). They show constancy in certain

8 characters. particularly in the structure of pharynx. This group is also considered as a sister group of both Cephalobina and Rhabditina (Furst von Eleven. 2000). However.

Diplogastrina shoes some transition towards the '1*yy lenchida especially in the structure of stoma and phar\nx and it clearly differs from the other two suborders.

Cephalobina and Rhabditina in having a aide. roomy and spacious huccal cm it\. a strong valuate median bulb and a non-valuate posterior phar\ ngeal hulk.

Diplogastrids have a short g. aeration time and therefore. are able to react rapidly on

fluctuations in food availability. When the microbial activity decreases. many form

inactive resting stages, called dauer larvae.

Faxonomic %\ork on the diplogastrid nematodes was also almost

simultaneously going on together with that of rhabditids. Schultz in Carus (1857)

described the genus Diplogaster. de Man (1876) described 7 vlopharvnx and

established the genera 1)iploga.vieroides and Odontohharvnzr (1912a, 1912b). He

i plated ()/ontophan-nr longieaToch10 from a rotting hyacinth bulb. Steiner (1914)

described the genus Demcneiel/a with D. cibourgensis as its type species ithout the

report of its male. f low ever. males were later reported by Paesler (1946) and `1e\ l

(1956). Micolet7.k,, (1922) placed (*/ontohhanvnv tinder Odontopharynginae, which

Steiner (1929) later on raised to family rank. Cobb (1920) established the genus

M%'ctolaimus and in 1924. he described the genus Neodiplogaster which has a

characteristic stomata) morphology. Rahm (1928) proposed Mononchoides with .tt

longicauda as its type species. Goodey (1929) proposed the genus Butlerius with B.

butleri as its type species.

Sachs (1950) gave some information on biology and ecology of free-living

nematodes especially rhabditids, diplogastrids and cephalobids and described

Diplogaster heur)cbu)c- (= Koerneria henrichyle), 1)iploga.ster erlcnlgensis (=

I\'aerak'rio t'rIan,s,'c'Tlsic). 1)ipin,L,aster lirsc'hun(m ae (= Koerneria birsehl,aaaue).

0 Diplogaster schwenrmlei (_ _lIononchoides sclnn-enimlei), Diplogaster zurstrasseni (_

Scichsia :urstrasseni) and redescribed several other species belonging to this group.

Paramonov (1952) described five new genera vi_.. Pareucliplogaster. Fictor.

Oigolaimella, Parc)i,golaimella and Dil)logasteritus and proposed the subfamily

Neodiplogastrinae. Ki rncr (195 4) described and illustrated several species vi_..

Dip/oga.ster inclrrs'u.s (= Anerneria nlc'II!ta). /)il)lo,raster lucclrri (= K. lucani).

Diplogu.rier sinodenc/rrmi (- K. .1inodendroni). Dih1(gaster (Eudil)loga.s'ter) goffar/i

(= K. gaffarti). I)iplogm-ter stvstenoceri (— K. rv.vtenoceri). Diplogaster ltr_iae (= K. luziae), Diplogaster sp/endichrs (= Afononchoide.s sl)lendidus), Diplogcisteroides

(ru.y~us, ;sTeodip/ogaster re /ala. iV. wacheki and Pseudodih/ogasteroides compositils.

\Veingcrtner (1955) classified diplogastrid nematodes into nine groups on the

basis of variations in the mouth cavity viz., i) grad/is group, ii) schneideri group, iii) nudicapitatus group. iv) rira/is group. v) inaequidens group. vi) coprophagus group,

vii) Iheritieri group. viii) levic%luus group and ix) strictus group. She (i.e.) also described Diplogaster dcn(Iro/)hilus (_ Ac1mstiehrs (lenc/rophi/us), D. microcercus

(= Paroigohinic110 micrrura). 1). lciick°Httl.s (= Fictor levir1ntns), D. lzi.stophoius (-

Knornerh hi.stohhora). D. lehto.sfnicrrlum (= Alononchoide.s Ieptospieu/u,i), D.

pvlophilu.s (= Al. pv/ophilus), D. elogans (= Al. elegans) and redescribed several other

species of this group.

AndrSssy (1958) described Diplogaster lepidus and provided keys for the

species with unpaired ovary. Timrn (1963) redescrihed D. cibou gen.si.s (Steiner.

1914). Goodey (196 3) raised Diplogasteridae to Diplogasteroidea and

Pseudod iplogasteroid idae to Pseudodiplogasteroidoidea and placed them under

Rhabditina. lie placed the subfamilies Diplogasterinae. Odontopharynginae and

I'v lopharynginae under the Tamil\ Diplogasteridae and svnonvmised the suhfamilies

Cephalobiinae Filipjev, 1934, Diplogasteroidinae Filipjev & Schuurmans Stekhoven,

10 1941, Demaniellinae Paramonov, 1951 and Neodiplogasterinae Paramonov, 1952

with Diplogasterinae Micoletzky, 1922. Under the subfamily Diplogastrinae he accepted 22 heterogeneous genera of which three viz., Rhabdiro(airnus Fuchs, 1931,

Mesodiplogaster Weingartner. 1955 and Micoletzkva Mcingartner, 1955 were

upgraded to generic rank and one new genus Hemidiplogasrer was proposed and

placed one genus each under Odontopharvnginae and Tylopharvnginae. Under the family Cylindrocorporidae Goudey, 1939 he accepted three genera viz.,

Cylindrocorpus Goodey, 1939, Goodeyus Chitwood, 1933 and Myctolaimus Cobb,

1920. Under Pseudodiplogasteroidoidea he accepted Pseudodiplogasteroides Kbmer,

1954 and Protodiplogasferoides (Ruhm, 1956) Paramonov, 1957.

Later, Taylor (1964) described Butlerius monhyslera having a single anterior gonad. In 1966, Kakuliya described Micoletzkya cordovector. Pillai and Taylor (1968) described Demaniella basili with some observations on its biology, isolated from sewage treatment plant at Urbana, Illinois and gave emendation of the genus

Demaniella. Goodrich, Hechler and Taylor (1968) described ,tfonondroide.s chatgi and M bollingeri from a waste water treatment plant. Pillai and Taylor (1968) described Bulletins micans collected from Illinois, with observations on its feeding habits. They also provided a key to the species of Butler/us. Fedorko and Stanuszek

(1971) described a new species, Pristionchus unforniis as a facultative parasite of

Leprinorarsa decendineara and Me/olDHtha melolontha from Poland. Calaway &

Tarjan (1973) analysed the stoma of Mortochoides Rahm, 1928 and described its structure and movements in feeding. Pokrovskaya (1974) redescribed Diplogastrellus mikuschi. Chin (1975) described the variation in the shape of the tail in

Cylindrocorpu.s curzii. Korenchenko (1975) isolated and described Cylindrocorpus vitauiasi from the tunnels of Ips subelongatus Morsch & ifylobius albosparsus.

11 Andrassy (1976) raised Cylindrocorporidae to superfamily rank and placed it

together with Diplogastroidea under Diplogastrina. but he did not accept

Pseudodipogasteroidoidea and placed Pseudodiplogasteroididae under

Diplogastroidea together with Odontopharyngidae Micoletzky, 1922,

Diplogasteroididae Filipjev & Schuurmans Stekhoven, 1941 and Diplogastridae

Micoletzky, 1922. He (I.e.) accepted the subfamilies Diplogastrinae, Dcmaniellinae,

Heteropleuronematinae and Tylopharynginae and proposed a new subfamily

Goffartinae under Diplogasteroididae and Paroigolaimellinae and Mononchoidinae (_

Neodiplogasteroidinae) under Diplogasteridae.

In 1978, Blinova el al. described Micoletzkya sexdentati ecolog ically and morphologically and also provided a toxonomic analysis of the genus Micoletzkya

Weingartner, 1955. Grootaert and Jaques (1979) described Butlerius degrissei. Zuilini

& Loof (1980) provided important notes on some species of Diplogastridae. Hunt

(1980) described Butlerius macrospiculum and Cylindrocorpus walkeri from St.

Lucia, West Indies. He extracted Butlerius macrospiculum from frass of Passalus sp.

(Coleoptera: Passalidae) and Cylindrocorpus walkeri was found to be an ecto-phoretic associate of the cerambycid beetle Lagochirus araneiformis (L.). Tsalolichin (1980) described and illustrated Butlerius gagarini and revised the genus Butlerius Goodey,

1929. He discussed eight species of Butlerius and provided their measurements and keys.

Inglis (1983) regarded the diplogastrid nematodes as a separate order,

Diplogastrida and divided them into five suborders: Diplogastrina, Alaninematina,

Cephalobina. Rhabdiasina and Strongyloidina, most of which included parasitic forms. In the same year, Geraert described and figured Afesodiplogaster pseudolheritieri.

Z Andrassy (1984) established Monobutlerius for three monodelphic species and also synonymised Butleroides Lordello & Zamith. 1957 and Butleriellus Meyl, 1960

with Butlerius. Andra'ssy (1984) considered Oigolaimella winchesi and O. asivmmetrica as O. agilis (Skwarra, 1921). Giblin and Kaya (1984) established

Aduncospiculum with A. halicti as its type species. It was found to be associated with a bee of the genus Halictus. Dassonville and Heyns (1984) described Mononchoides aquaticus and M. grad/is from fresh water from South Africa.

Again, Andrassy (1984) in his exhaustive account on class Nematoda divided

Diplogastrina into three superfamilies — Cylindrocorporoidea Goodey, 1939,

Odontopharyngoidea Micoletzky, 1922 and Diplogastroidea Micoletzky, 1922. In

Cvlindrocorporoidea Goodey, 1939 he erected the family Cylindrocorporidae

Goodey, 1939 and placed Myctolaimus, Goodeyus, Protocylindrocorpus (Rahm,

1959) Paramonov, 1964, and the newly proposed genus Myctolaimellus under it. He

(l.c.) upgraded the family Odontopharyngidae Micoletzky, 1922 to superfamily

Odontopharyngoidea Micoletzky, 1922 and placed OdontopharynY De Man, 1912 and proposed Zullinius. Diplogastroidea was divided into six families —

Pseudodiplogasteroididae Korner, 1954. Diplogasteroididae Filipjev & Schuurmans

Stekhoven, 1941, Diplogastridae Micoletzky. 1922, Neodiplogastridae Paramonov,

1952, Tylopharyngidae Filipjev. 1934 and upgraded Heteropleuronematinae

Andrassy, 1970 to family Heteropleuronematidae Andrassy, 1970. Under

Pseudodiplogasteroididae he placed Pseudodiplogasieroides Korner, 1954 and

Protodiplogasteroides (Ruhm, 1956) Paramonov, 1957. Under Diplogasteroididae he placed Diplogasteroidinae Filipjev & Schuurmans Stekhoven. 1941 for

Diplogasteroides De Man, 1912, Paramonovnema Andrassy, 1968 and Goffartia

Hirschmann, 1952 and proposed Rhabditolaiminae for Rhabditolaimus Fuchs, 1915,

Rhabditidoides Rahm, 1928, Rhabdontolaimus (Fuchs, 1931) Filipjev & Schuurmans

13 Stekhoven, 1941 and newly proposed Fuchsnema. Diplogastridae was further

classified into three subfamilies — Demaniellinae Paramonov, 1951, Diplogastrinae

Micoletzky, 1922 and Paroigolaimellinae Andr3ssy, 1976. Under Demaniellinae he

included Metadiplogaster Meyl, 1961 and Demanieia Steiner, 1914. Diplogastrinae

included Acrostichus Rahm, 1928, Diplogasteritus Paramonov, 1952, Afasseyus

Paramonov, 1964, Allodiplogaster Paramonov & Sobolev in Skrjabin, Shikhobalova,

Sobolev, Paramonov & Sudarikov, 1954, Diplogasteriana Meyl, 1961, Butlerius

Goodey, 1929. Mesodiplogastcroides Khera, 1969, Anchidiplogaster Paramonov,

1952, Diplogaster Schultze in Carus, 1857, Gerthornus Massey, 1966 and proposed

Monobutlerius under this taxon. The subfamily Paroigolailmellinae included

Eudiplogasterium Meyl, 1961 and Paroigolaimella Paramonov, 1952. Under

Neodiplogastridae was placed Neodiplogastrinae Paramonov, 1952 for Pristionchus

Kreis. 1932, Oigolaimella Paramonov, 1952, Fietor Paramonov, 1952, Micoletzkya

(Weing9rtner, 1955) Paesler. 1962, Mononchoides Rahm, 1928, Diplenteron

Andressy, 1964, Koerneria Meyl, 1961 and A-'eodiplogaster Cobb, 1924 while,

Glauxinematinae was newly proposed for Pareudiplogaster Paramonov, 1952 and

Glauxinema Allgdn, 1947. The last two families of this superfamily viz.,

Heteropleuronematidae and Tylopharyngidae included one genus each viz.,

1-feterapleuronema Andrassy, 1970 and Tylopharynx De Man, 1876 respectively.

Ebsary (1986a) described Butlerius canadensis and porposed the genus

Parabutlerius for the monodelphie species. Ebsary (1986b) also described and illustrated Mononchoides andersoni, Koerneria terranovus and Koerneria mulveyi from Canadian localities. Sudhaus & Rehfeld (1990) described Diplogaster coprophilus (= Paroigolaimella coprophilus), D. bernensis (= P, bernensis) and D. aflinis (= P. n{Tnis) keeping all the diplogastrid genera as subgenera of Dtplogaster.

Chitambar (1990, 1995) described new predaceous nematode. ifonohutleriue

14 macrogubernculum extracted from soil around roots of Pathos sp. growing in a

nursery in Santa Barbara, California, U.S.A. and also redescrihed Oigolaimella

carinata (Zullini, 1981) collected from polluted water in Italy. The controversy

regarding origin and relationships of the genus Tylopharync (Andrassy, 1984;

Maggenti, 1963, 1983; Poinar, 1983; Siddiqi, 1980) was resolved by De Ley et at

(1993) when they suggested that there is no strong affinity between the stoma of

Tylopharynx and the stylet of Tylenchida, based on detailed observations of the

ultrastructure of the stoma stmeture of T. foetida. Andrassy (1984) placed it in the

family Tylopharyngidac. In 1995, Gagarin reviewed the genera Ficlor and Koerneria

and added a new species Fictor shoshini to the genus Fictor. Kuhne (1995) described

Diplogaster labiomorphus, phoretically associated with a dung beetle and compared it

with D. henrichae Sachs, 1950 and D. hirschmannae Sachs, 1950 morphologically.

Morand and Baker (1995) established Hugotdipingaster with H. neozealandia as its

type species. They found it to parasitise an endemic slug, Anthoracophorus

bitentaculatus in New. Zealand. Kiontke and Sudhaus (1996) revised the subgenus

Diplogastrellus and added a new species, Diplogastrellus cerea isolated from

decaying cactus in Arizona, USA and commented on the phylogenetic relationships of

the twelve species in this subgenus and provided stale and female keys to species and also provided information on their habitats and geographic distribution. They synonymized Metadipiogaster with Diplogastrellus mainly on the basis of location of nerve ring, shape of gubemaculum and arrangement of caudal papillae. Sommer no!.

(1996) described Pristionchus paciJicus morphologically as well as at molecular level.

Gagarin (1998a) added one more species to the genus Fictor. F tumidus collected from rotten vegetables. In the same year (I 998b) he gave the review of the genus

Mononchoides and described the two new species namely, Al. paramonovi and M microstomus from Russia, and gave measurements and differential keys to 24 valid

15 species of this genus and also considered Diplogaster (Eudiplogaster) eleganr and D.

(E) leptospiculum as species incertae sedis.

Furst von Lieven & Sudhaus (2000) accepted Diplogastrina and studied

functional morphology of the buccal cavity and provided a first outline of the

phylogenetic relationships of the suborder. They also confirmed that Tylopharync and

Mononchoides are closely related. However. Furst von Lieven (2001, 2002a)

reconstructed a cladogram of the diplogastrids based on the intensive study, mostly of

the stoma using light microscopy and SEM. In 2001, Wu et al. described a new

species Tylopharynx clariantphida and redescribed Tylopharynx foefida (BUtschli,

1874) Goffart, 1930. Kointke et al. (2001) gave a detailed morphological description

of Diplogasteroides nasuensis Takaki, 1941 and D. magnus Volk, 1950 using LM and

SEM. Dauer larvae of both the species were found on the wood cockchafer

(Melolontha hippocastam) larvae and adults, but on adult beetles, dauer larvae of D.

nasuensis entered the genital pouch. whereas those of the D. magnus were found

under the hind coxae. Furst von Lieven (2002b) considered Tylopharynx de Man,

1876 to be the sister taxon of the genus Neodsplogaster Cobb, 1924. These two genera

share a narrow and elongated cylindrical stegostom extending to the periradial struts

which possess a subdorsal apodeme as a synapomorphic character.

De Ley and Blaxter (2002, 2003) placed these nematodes under the name

Diplogasteromorpha, an infraorder under the suborder Rhabditina. They grouped the eight families under the three superfamilies- Diplogastroidea, Cylindrocorporoidea and Odontopharyngoidea.

Poinar et at (2003) included five genera into the family Cvlindrocorporidae

Goodey, 1939 on the basis of male bursa and female gonads; Cylindrocorpu.s Goodey,

1939; Myctolaimus Cobb, 1920; Mycloloimellus Andrassv, 1984;

Prolocylindrocorpus Ruhnt, 1959; Goodeyus Chitwood, 1933. Furst von Lieven

16 (2003a) revised the genus Ogolaimella and added two new species to the genus O.

kruegeri and O. nine. Besides describing C:ylindrocorpus inevectus, Poinar et al.

(2003) provided a key to the genera of Cylindrocorporidae and proposed a new family

Longibuccidae for the members of the genus Longibucca, previously placed in

Cylindrocorporidae.

Sudhaus & FUrst von Lieven (2003) in their cladistic approach of classifying

diplogastrid nematodes, synonymized several genera and provided a more reliable

classification of diplogastrids based on their phylogenetic relationships and also

provided a catalogue having a list of genus group names and their synonyms and a list

of all species, with information on their habitats. Three genera were synonymized

with Acrostichus; five genera with Butlerius. No synonymy was suggested for both

Cephalobium Cobb, 1920 and Demaniella and Diplogaster and also for

Diplogasteriana. With Diplogasteroides they synonymized six genera. With

Diplogastrellus they synonymized two genera and with Fictor they synonymized four

genera. Only one genus was synonymized with Goffartia. For Heteropleuroncma

Andrassy, 1970, Hugotdiplogaster Morand & Barker, 1995, Longibucca Chitwood,

1933 and Afehdinema Farooqui, 1967 too they suggested no synonymy. With

Koerneria four genera and with Aficole: ya two genera were synonvrnized. With

Mononchoides seven genera were synonoymized and with Afyctolaimus five genera

were synonymized. No synonymy was suggested both for Areodiplogaster and

Parasiiodiplogaster. Only one genus each was synonymized with Odontopharvnx,

and Oigolaimella. Two genera were synonymized with Paroigolaimella. Seven

genera were synonymized with Pri.ctionchus. One genus each was synonymized with

Pseudocliplogasteroides. Rhabditidoides and Tvlopharynr. For .Sachsia Meyl. 1960 no

synonymy was suggested. Thus, in all, they accepted only 28 genera of the published

83 genera and listed 312 nominal species and kept all under one family

17 Diplogastridae. Of the 28 genera, five genera viz., Heteropleuronema, Sachsia,

Parasitodiplogaster, Cephalobium and Hugotdiplogaster were kept as genera of

uncertain position.

Recently, after the review of Sudhaus & Ftlrst von Lieven (2003), Kanzaki &

Futai (2004) isolated a new species Cylindrocorpus anoplophorae from genital

capsule of the white-spotted longicorn beetle, Anaplophora malasiana.

Recently, Andrassy (2005) placed five suborders under the order Rhabditida-

Teratocephalina, Cephalobina, Myolaimina, Rhabditina and Diplogastrina. Under

Diplogastrina he distinguished nine families assigned to three superfamilies: 1.

Cylindrocorporoidea Goodey, 1939, 2. Odontopharyngoidea Micoletzky, 1922 and 3.

Diplogastroidea Micoletzky, 1922. Two families- Cylindrocorporidae Goodey. 1939

(four genera: Cylindrocorpus Goodey, 1939, Goodeyus Chitwood, 1933, Myctolaimus

Cobb, 1920 and Protocvlindrocorpus RUhm. 1959) and Longibuccidae Poinar,

Jackson, Bell & Wahid, 2003 (One genus: Longibucca Chitwood, 1933) were placed

under the superfamily Cylindrocorporoidea Goodey, 1939. One family-

Odontopharyngidae Micoletzky, 1922 (one genus: Odoniopharynx, de Man, 1912)

was placed under the superfamily Odontopharyngoidea Micoletzky. 1922. Six

families- Pseudodiplogasteroididae Korner. 1954 (one genus: Pseudodiplogasteroides

Komer, 1954); Diplogasteroididae Filipjev & Schuurtnans Stekhoven, 1941 (eight

genera: Demaniella Steiner, 1914, Diplogasteroides de Man, 1912, Fuchsnema

Andrdssy, 1984, Goffartia Hirschmann. 1952, Hugotdiplogarter Morand & Barker,

1995, Mehdinenta Farooqi, 1967, R/rabditidoides Rahm. 1928 and Rhanditolaintus

Fuchs. 1915): Diplogastridae. Micoletzky, 1922 (thirteen genera: Acrostichus Rahm,

1928, Anchidiplogasver Paramonov, 1952, Butlerius Goodey, 1929, Cephalohiunr

Cobb, 1920, Costanemella Andrassy, 2001, Diplogaster Schultze in Carus, 1857,

Diplogasteriana Meyl, 1961, Diplogasieritus Paramonov, 1952, Eudiplogasterium

18 Meyl, 1961, Monobutlerius Andrassy. 1984, Parasitodiplogaster Poinar, 1979,

Paroigolaimella Paramonov, 1952, Peterngus Ahmed, Abroad & Mahamood, 2004);

Neodiplogastridae Paramonov, 1952 (ten genera: Diplenteron Andrassy, 1964, Fictor

Paramonov. 1952, Glouzinema Allgen, 1947, Koerneria Meyl, 1961, Micoletzkya

Weingimner, 1955, Mononchoides Rabin, 1928, Neodiplogaster Cobb, 1924,

Oigolaimella Paramonov, 1952, Pareudiplogaster Paramonov, 1952 and Pristionchus

Kreis, 1932); Heteropleuronematidae Andrassy, 1970 (one genus: Heleropleuronema

Andr3ssy, 1970); Tylopharyngidae Filipjev, 1934 (one genus: Tylopharynx Filipjev,

1934) were placed under the superfamily Diplogastroidea Micoletzky, 1922.

However, Andrassy (2005) did not accept the synonymy of Mononchoides

with Glauxinema based on the presence of a left subventral plate in the buccal cavity

of Mononchoides species and accepted only 29 valid species of Mononchoides and 9

valid species of Glauxinema. Kanzaki et at. (2006) isolated and described a new

species of Myetofaimus Cobb, 1920 M platypi from the underside of the elytra of oak

borer beetle, Platypus quercivorus Murayama. Poinar et al. (2006) described

Chroniodiplogaster formosiana collected from a Chinese termite Odontoterines formosanus Shiraki (Isoptera: Termitidae). Giblin-Davis et al. (2006) described

Parasitodiplogaster laevigara recovered as dauer (infective) juveniles from the

hemocoel of newly emerged adult females of the fig wasp Pegoscapus sp.

(Agaunidae). They also reported molecular phylogenetic comparisons of two expansion segments (D2, D3) of the large subunit (LSU) rRNA gene from

Parasitodiplogaster loevigato with several species of Parasitadiplogaster from

Panama.

Kanzaki et I. (2008) described and illustrated two species of Neodip(ogaster, namely, N. crenatae isolated from bark beetles Scolycoplatypus daimio and S. shogun

(Scolytidae) and N. acaloleptae isolated from the cerambycid beetle Acalolepta

19 lururiosa. Furst von Lieven (2008) described a hermaphrodite diplogastrid, Koerneria

sudhausi and its egg shell formation from zygote and uterine components. Kanzaki et

al. (2008) described Diplogastrellus metamasius isolated from a West Indian

sugarcane weevil, Metamasius hemipterus (L.) from southern Florida and Costa Rica.

They cultured this species from dauer juviniles recovered during dissection of the

West Indian sugarcane weevil, Metamasius hemipterus (L.) (Cleoptera:

Drvophthoridae) which showed a phoretic association with M. hemipterus. Furst von

Lieven & Sudhaus (2008) described a new species, Oigolaimella attenuata which was

found to be associated with the colonies of termites belonging to the genus

Reticulitermes from Corsica (France) and USA. The new species was found in the

preoral cavities of the termites for internal phoresis and was found to be associated

with non-pathogenic gut-inhabiting flagellates of the taxon Kinetoplastida. They also described its life cycle and development including spermiogenesis, sperm transfer, sperm competition and fertilization in detail. Sudhaus (2008) also discussed the evolution of insect parasitism in Rhabditid and Diplogastrid nematodes.

Kanzaki et al. (2009) established a new genus Teratodiplogaster under the family Diplogastridae. isolated from the Syconia of Ficus raceniose in Queensland and Western Australia with T. fignewnruni as its type species. Bartholomaeus et al.

(2009) isolated and described Para.sitodiplogaster australis from Ficus virens moraceae in Australia. Kanzaki et al. (2009, 2010a, 2010b) isolated a few species of

Acrostichus, A. rhynchophori from the palmetto weevil, Rhynchophorus cruentatus

Fabricius (Coleoptera: Curculionidae), in southern Florida and palm weevil. R. palmarum L., in Colombia. Costa Rica and Trinidad, .4crovtichus purl from the body of a sweat bee, Augochlora purer nrvsieri and Acrostichus megaloptae from the

Dufour's gland of female bees and aedeagus of male bees. Megalopia spp. in Central

America.

20 Recently, Hodda (2011) accepted the order Diplogastrida Inglis, 1983 and

placed it under the superorder Rhabditica Hodda, 2007. He divided the order

Diplogastrida into following suborders: Chambersiellina Hodda, 2007; Diplogastrina

Paramonov, 1952 and Myolaimina Inglis, 1983. One superfamily, one family, two

subfamilies, seven genera and sixteen species were placed in the suborder

Chambersiellina Hodda, 2007. The suborder Diplogastrina was further divided into

two superfamilies: CyIindreorporoidea T. Goodey, 1939 (Andrassy, 1976) and

Diplogasteroidea Micoletzky, 1922 (Goodey. 1963). Two families, namely

Cylindrocorporidae T. Goodey. 1939 (3 genera, 22 species) and Odontopharyngidae

Micoletzky, 1922 (1 genus, 2 species) were placed in the superfamily

Cylindrcorporoidea. Under the superfamily Diplogasteroidea, six families were

placed, viz. Cephalobiidae Travassos & Kloss, 1960 (2 subfamilies, 6 genera, 30 species): Diplogasteridae Micoletzky, 1922 (Steiner, 1929) (1 subfamily, 15 genera,

239 species); Diplogasteroididae Paramonov, 1952 (3 genera, 40 species);

Neodiplogastridac Paramonov, 1952 (Andrassy. 1984) (1 subfamily, 2 genera, 10 species); Pseudodiplogasteroididae De Ley & Blaxter, 2002 (1 genus, 2 species);

Tylopharyngidae Filipjev, 1918 (I subfamily, 2 genera, 3 species). The suborder

Myolaimina Inglis, 1983 was further splitted into two superfamities:

Carabonematoidea Stammer & Wachek, 1952 (Hodda, 2007) (1 family, I genus, I species) and Myolaimoidea Goodey, 1963 (Hodda, 2007) (1 family, 4 genera, 22 species).

Steel et al. (201 1) described 1llononchoides composticola isolated from compost. Furst von Lieven et al. (201 1) introduced a genus Cutidiplogaster with C. manati isolated from skin lesions of manatees, Trichechus manarus manutu.r, kept in

Okinawa Churaumi Aquarium, Japan. Kanzaki et al. (2012a) described and illustrated two new species, namely, Teratodiplogaster martini and Parasitotliplogaster

21 doliostoma from Africa. Kanzaki el al. (2012b) also described two new species of

Pri.stionchus P. fissidenraru.s from Nepal and La Reunion Island and P. elegans from

Japan.

For years, morphological identification was the only method used to identify nematodes. As our knowledge of nematodes of agronomical importance increased it became clear that morphology alone did not reveal the complete picture of observed pathological differences between populations within morphologically delimited species. Thus, new methods have been looked for that could better predict observed pathological behaviours among populations within species. New approaches are coming in use to aid species identifications within the context of a classical morphological system. Currently a shift from the purely phenotypic to using a combination of both phenotypic and molecular methods is observed (Powers et al.,

1997; Powers, 2004). Nowadays. phylogenetic species concept has gained more support (Adams, 1998, 2002) and ways to extend its theoretical appeai into practicality have been evaluated (Nadler, 2002). An evolutionary framework is necessary to understand the genera and their relationship to other groups and for the designation and verification of new species. Although morphology and life history prove useful, gene sequences provide an independent and alternative data set for phylogenetic inferences. Thus, in addition to the ultrastructural (scanning electron microscopic and transmission electron microscopic observations) and taxonomic descriptions molecular phylogeny proves useful. Numerous molecular techniques have been developed that are capable of identifying and quantifying nematodes at the species level and below. Techniques such as isozyme pattern analysis, restriction fragment length pelymorphism (RELP) analysis, random amplified polymorphic

DNA (RAPD) analysis, polymerase chain reaction (PCR), quantitative polymerase chain reaction (qPCR) and sequencing of diagnostic rDNA regions have all been used

22 successfully to identify and quantify several agriculturally important plant parasitic

nematodes. These methods have their own advantages and limitations. DNA

sequences of marker genes, Denaturing Gradient Gel Electrophoresis (DGGE) and the

more recently developed method of pyrosequencing are the three methods employed

in biodiversity studies of free-living forms. Andre et al. (2002) highlighted the need

for the development and consistency of methods in soil faunal monitoring;

commenting that molecular techniques for community analysis are now widely used

in soil microbiology and have greatly expanded our knowledge of soil nematodes.

Thus, molecular methods provide an alternative to the traditional morphological

identification for routine assessment of described species. Their application has enabled profiling of environmental samples of soil microbial populations, overcoming the need to culture and identify bacteria and fungi from complexes mixtures (Amann et al., 1995) and may reduce the taxonomic expertise currently required to characterise microfaunal communities.

Analysis of allozyme patterns was the first biochemical method to characterize and diagnose nematode species. Random amplified polymorphic DNA (RAPD) anaiNsis has been used to assess genetic variation among isolates of the same species

(Dona et al., 2001; Cofcewicz et al., 2005; Vieira et al., 2007; Devran etal., 2009) and between species (Cofcewicz et al., 2005). Bandi et al. (1995) compared patterns generated using RAPD with those of allozymes. DGGE has been shown to underestimate nematode biodiversity (Foucher et al., 2004; Cook et al., 2005). But, even with this limitation, it can be useful to evaluate patterns of nematode diversity over time (Wang et al.. 2008). Multiplex PCR has been applied in many areas of

DNA testing, including species identification, analyses of deletions, mutations, polymorphism. quant itative assays, and reverse transcription PCR (Elnifro et al.,

2000; Subbotin and Moens. 2006). Amplification and sequencing of diagnostic

23 regions of nematode DNA have become the major source of new information for

advancing our understanding of evolutionary and genetic relationships (Hajibabaei et

al., 2007; Meldal etal., 2007). With regard to free living nematodes, DNA sequencing

has been used for rapid and accurate taxonomic identification and understanding

population genetic structure in marine nematodes (De Ley et al., 2005; Bhadury et al.,

2006; Derycke el al,, 2007. 2008;).

The use of these methods will encourage the inclusion of this diverse animal

group in biodiversity and environmental studies. Such an inclusion would be an

important step towards a better understanding of the biology and ecology of nematodes. Therefore, molecular methods will help us realize our goals, i.e., to inventory and understand the biological diversity and ecological role of nematodes.

Nonetheless, despite some difficulty associated with the use of morphology in the taxonomy of some groups of nematodes, the extensive amount of information accrued on various aspects of nematode biology is currently linked with morphology.

Consequently, this makes the integration of morphology in taxonomic studies relevant and essential.

Blaxter et al. (1998) proposed the first molecular phylogenetic framework of the phylum nematode. They constructed a database of small subunit (SSU) sequences from 53 taxa, including 41 new species to present a phylogenetic tree of nematodes.

Sequences were aligned with reference to a secondary structure model and on the basis of similarity. They recognized three major clades: Glade I groups the vertebrate- parasitic order Trichocephalida with the insect-parasitic Mermithida, plant-parasitic

Dorylaimida and free-living Mononchida. Clade II links the plant-parasitic

Triplonchida with the free-living Enopllda & Monhysterida and Glade C+S groups

Chromodorida and Secernentia. Within Seccrnentia three major clades were identified

(Glade III, IV and V). Clade III represents a grouping of vertebrate- and arthropod-

24 parasitic taxa . from the orders Ascaridida, Spirurida, Oxyurida and Rhigonematida.

Clade IV groups the plant parasitic orders Tylenchida and Aphelenchida, the

vertebrate-parasitic genus StronD,loides and the entomopathogenic genus

Steinernema with free-living bacteriovores of the rhabditid families Cephalobidae and

Panagrolaimidae. Clade V agroups C. elegans and other members of the suborder

Rhabditina with the vertebrate-parasitic order Strongylida, the entomopathogenic

genus Heterorhabditis and the order Diplogastrida.

An accurate phylogeny for nematodes including their relationship to other animals has major consequences for our understanding of how diversity has evolved

in morphology, genes, developmental mechanisms, behavior, and life history, but also

for studies of comparative functional genomics. Reid et al. (1997) presented molecular taxonomy and phylogeny of entomopathogenic nematode, Steinernema

Travassos, 1927 by RFLP analysis of the ITS region of the ribosomal DNA repeat unit. Dorris et al. (1999) presented molecular analysis of nematode diversity and discussed the evolution of parasitism. Liu el al. (1999) presented molecular differentiation and phylogeny of entomopathogenic nematodes based on ND4 gene sequence of mitochondrial DNA from 15 heterorhabditid nematode isolates, representing 5 species collected from different regions of the world, by using polymerase chain reaction (PCR) and direct-sequencing of PCR products.

Floyd et al. (2002) developed a molecular operational taxonomic unit

(MOTU) scheme for soil nematodes using a molecular barcode, derived from single- specimen polymerase chain reaction (PCR) and sequencing of the 5' segment of the small subunit ribosomal RNA (SSU) gene. A total of 23 MOTU were defined from only 240 sequences. [Many MOTU could readily be assigned to classical, morphologically defined taxonomic units using a database of SSU sequences from named nematode species. The MOTU technique allows a rapid assessment of

25 nematode taxon diversity in soils. Correlation with a database of sequences from

known species offers a route to application of the technique in ecological surveys

addressing biological as well as genetic diversity.

De Ley and Blaxter (2002, 2004) updated the classification of the phylum

Nematoda using molecular data available from additional species, with morphological

data to assist the placement of taxa for which SSU sequences were not yet available.

They used SSU phylogenies to develop a novel classification reflecting recent

evolutionary findings and proposing the infraorders Cephalobomorpha,

Panagrolaimomorpha and Tylenchomorpha, all within a considerably expanded

suborder Tylenchina. Faucher et al. (2004) assessed nematode biodiversity using

DGGE of IRS rDNA following extraction of nematodes from soil.

Pires-da Silva and Sommer, 2004; Rudel et al., 2005; Zheng el al., 2005

studied the developmental, genetic and molecular analysis of sex determination and

vulva and gonad formation in Pristionchus pacificus. The hermaphroditic nematode

Prisnonchus pac fcus Summer, Carla, Kim and Sternberg, 1996 has been developed

as a satellite organism in evolutionary developmental biology (Hong and Sommer,

2006). Herrmann et al. (2006a, b) found that the Pristionchus live in close association

with scarab beetles and the Colorado potato beetle (CPB) in western Europe and

eastern North America.

Bhadury et al. (2006) studied nuclear and initochondrial genomic regions and

evaluated their potential use for marine nematode identification. Their results showed

That I 8S ribosomal RNA gene amplified most reliably from a range of taxa and can be

useful for rapid molecular assignment of unknown nematode species. Stock and

Nadler (2006) revised the taxonomic status of several species from the genus

Panagrellur combining classical morphological data and molecular sequences and presented its phylogenetic relationships. Mayer et al. (2007) presented a phylogenctic

26 framework of the genus Pristioncbus to interpret the ecological patterns. Herrmann et

al. (2007) described 40 isolates of P. pacificus from scarab beetles in Japan, in par-

ticular from the oriental beetle Exonrala (Anomalaa oriema1is. Mayer et al. (2007)

provided the phylogeny of all 18 availahle Pristionchus species based an a set of 27

ribosomal protein genes encompassing a total of 10,971 bp. The phylogenetic tree

provides evidence for North American and European clades, which are embedded in a

deeper Glade that includes Asian species. Of the 18 Pristionchus species, 13 are

gonochoristic and five are hermaphroditic. The phylogeny indicated that all

hermaphroditic species have arisen independently within the genus Pristionchus.

Meldal et al. (2007) revised the molecular phylogeny of De Ley and Blaxter (2002)

by adding sequences to the nematode SSU data set from previously under-represented

marine taxa and from additional terrestrial and parasitic groups. They added SSU rDNA sequences for 100 un-sequenced species of nematodes, including 46 marine taxa. Sequences for 200 iaxa have been analysed based on Bayesian inference and logDet-transformed distance analysis. The resulting phylogenies provide support for the re-classification of the secementea as the order Khabditida that derived from a common ancestor of chromadrean orders Acreolaimida. Chcmnadorida, Desmodorida,

Desmoscolecida, and Monhysterida. Their analysis also support the position of

Bunonema close to the Diplogasteroidea in the Rhabditina. They also proved that SSU rDNA genes are very effective in the recovery of many monophyletic group within the phylum Nematoda and provided clarification of relationships that were uncertain and controversial. Sudhaus and Kiontke (2007) studied the genus Caenorhabdirit extensively and proposed the nominal species Caenorhabditis brenneri primarily based on "cross-breeding experiments, biogeography and DNA sequences".

Mayer et al. (2009) established a molecular phylogeny of beetle-associated diplogastrid nematodes which suggested host switching rather than nematode-beetle

27 coevolution. Porazinska et al. (2009) reported the use of high-throughput sequencing

for metagenomic analysis of nematode diversity using coding sections of the small

and large subunit of rRNA genes.

Recently. pyrosequencing has emerged as a new sequencing technology (Creer

et al., 2010). The most important applications of this technology is the ability to

identify large numbers of species from complex communities and recently it has been

widely used to generate the genome sequences of complex environmental samples

(Edwards et al., 2006; Joseph el al., 2009). Roesch et al. (2007) employed the method

to investigate soil microbial diversity, and others have used it to study plant

pathogenic fungal communities (Das et al., 2008; Buee et al., 2009). In nematology, it

has been tested and has shown encouraging results for the analysis of nematode

diversity from metagenomic samples (Creer et al., 2010). Ababe et al. (2011)

presented a review of various techniques used in taxonomy of free-living and plant

parasitic nematodes and critique those methods in the context of recent developments

and trends including their implications in nematode taxonomy, biodiversity and

biogeography.

Accuracy of identification is fundamental to our understanding and communication of the ecological role of any organism. I lugot (2002) emphasized the need for correct identification and the role of taxonomy as a science. Biodiversitv can be assessed at any taxonomic level, and by either molecular or phenotypic characters or a combination of both.

In India, Nematology started as one of the disciplines of agricultural sciences.

Barber (1901) was the first to record the infestation of root-knot nematode on tea in

South India. The 1960s could be regarded as the most active phase for the growth of nematolog\ in general and nematode taxonomy in particular. Nematological researches in India have focussed more on the plant parasitic and animal parasitic

28 groups. No extensive work has so far been carried out on the taxonomy of free-living

nematodes. Only a few scientists have worked on this heterogeneous group during the

late 60s and 70s. One of the reports which is considered earliest is the establishment

of a new genus Tridontus by Khera (1965) collected from banks of still and running

waters under the subfamily Diplogasterinae, with T. longicaudatus (= Mononchoides

longicaudatus) as its type species. Khera (1969) described Mesodiplogasteroides and

in 1970 Paradoxogaster (= Anchidiplogaster) and Gobindonema filicaudatum (=

Koerneria filicaudata) without the report of its male in his description, from the banks

of a freshwater lake near Dehradun, Uttarakhand, India. Suryawanshi (1971)

described Tawdenema and Syedella which later on were synonimized with

Acrostichus and Pareudiplogaster respectively. Jairajpuri et al. (1973) redescribed

Tridontus longicaudatus (Mononchoides longicaudatus) and synonimized Syedella

with Tridontu.s. In the suborder Rhabditina, Khera (1969) described the genera

Operculorhabditis in Rhabditonematinae, Saprorhabditis in Rhabditonematinae and

Praeputirhanditis (= (J'uticularia) in Rhabditinac and in 1971 Paradoxorlrabditis in

Protorhabditinac. Huseni et al., (1997) described Bunonema irregularis, B. minutum and Pterygorhabditis superbus under the superfamily Bunonematoidea. Tahseen and

Jairajpuri (1988) reported Teratorhabditis andrassvi and also described its biology in detail.

Siddiqi. Bilgrami & Tabassum (2004) described Mononchoides gaugleri from agricultural farm. Hussain et al. (2004) were the first to describe two new species,

Myctolaimus neolongistoma and M. kishtwarensis, from India. Ahmad et al. (2004) established a new genus under the family Diplogastridae, I'eterngus from the fluid in the pitchers of?Vepenthes anrpullaria..V. grad/is and A". rafilesiana in Singapore. with

I'. nepenthi as its tN pe species. Ahmad et al. (2005) provided a detailed description of two species of the genus Diplogasuellus Paranionov, 1952, namely D. thoubalicus

Wi and D. heynsi (Kiontke & Sudhaus, 1996) Sudhaus and Ftlrst von Lieven, 2003 during

their survey of diplogastrid nematodes from India. Maharnood et at (2006a, 2006b)

added two new species to the genus Fietne Paramonov, 1952, namely F. denticulatus

and F selosus isolated from horse dung and farmyard manure respectively and one

species to the genus Demaniella Steiner, 1914, namely D. keirakensis. Mahamood et

at (2007) also described and illustrated Afononchoides megaonchus from decaying

banana rhizome. Rahmat er at (2008) described and illustrated Diplogatrellus grad/is

(Butschli, 1876) Paramonov, 1952, D. sikarar and Ficlor composticola using SEM

and LM details from India. Ahmad etal. (2009) redescribed Butlerius burleri Goodey,

1929 with additional information from the specimens collected in South Korea.

Mahamood and Ahmad (2009) described two new species, namely Diplogasteroides

uriceus and Rhabditidoides papilla/us collected from chicken manure and farmyard

manure respectively. Recently. Mahamood and Ahmad (2010) redescribed Koerneria

filicaudata (Khera, 1970) Andrassy, 1984 with the first report of its male.

Review on biology of free-living nematodes

The bulk of research work carried out in Nematology pertains primarily to

fundamental aspects like taxonomy and morphology. But an increasing interest in the

subject has given way to a greater diversification of research programmes leading to

investigations on ecology, biology, physiology, host-parasite relationships, behaviour, etc. of nematodes. Free-living species including nematodes that feed on bacteria, fungi, and other nematodes are abundant, yet the vast majority of species encountered are poorly understood biologically. Some pioneering works in this field have been done by workers like Klinger (1958, 1959, 1961, 1963, 1965, 1966 & 1970), Wallace

(1958-63, 1968-70), Croll (1965-67, 1970-72 & 1974-76), Bird (1959-62). Green

30 (1966, 1967), and others which aroused the interest of workers all over the world.

Bastian (1846), Butschli (1873) and rley (1886) observed feeding in nematodes.

Goette (1882) first described the embryology of Rhabditis. In 1899, Maupas

studied moulting and post-embryonic growth of larvae of Rhabditis (Choriorhabditis)

caussaneli and Rhabditis (Choriorhabditis) pellio. Chitwood and Chitwood, 1950

studied the ingestion in Rhabditis and other members of the superfamily

Rhabditoidea. Thorne (1961) suggested that food particles are broken down during

ingestion in Rhabditis (Cephaloboides) oxycerca and he attributed this to the

shredding action of the teeth at the base of the stoma.

The chemical and physical factors of the environment influences the behaviour

of nematodes. Any change in the environment brings about a corresponding change in

the activity of these animals. The nature of the change and the mechanism involved

forms the basics of behavioural studies. Earlier workers, Fraenkel and Gunn (1961) and Prinjle (1963) have distinguished and defined various kinds of responses particularly orientation and attraction and kinesis and taxes and established terminologies now commonly accepted. Doncaster, 1962 observed feeding mechanisms in Rhabditis oxycerca and Pelodera lambdiensis in detail. A substantial amount of work has been done on the locomotory behaviour by Wallace (1958-63,

1968), Gray (1953). Gray and Lissmann (1964). Crofton (1971), Croll (1971), and

Azmi and Jairajpuri (1976, 1977) and orientation to physical and chemical stimuli by

Klingler (1958, 1959, 1961 & 1963), Wallace (1968). Doncaster and Shepherd (1967) and Croll (1974) described the behaviour of the juveniles within the egg and the mechanism of hatching, while Hechler and Taylor (1966) and Chin and Taylor (1970) described the behavioural activities of the moulting stages of some plant-parasitic and free living species. The behaviour of males during mating was studied and described by Greet (1964). Jones (1966) and Chin and Taylor (1969).

31 Free living saprophagus nematodes have been extensively used as an

important tool in biochemical and nutritional studies (Dougherty et al., 1959,

Nicholas, 1959, Vanlleteren, 1973). Their relatively short life cycles and high

reproductive rate are an advantage in these studies. Chuang (1962) studied the

embryogenesis and post-embryogenesis of Riuebditis teres and Thomas (1965) gave

an account of the life cycle of Acrobele.s eomplexus. Greet (1964) noticed the

existence of mutual attraction in Panagrolaimus rigidus. Jones (1966) observed sex

attraction and copulation in Pelodera teres. Chin and Taylor (1969, 1970) and Chin

(1977) have made an excellent study of the biology of Cylindrocorpur sp. Some

aspects of the nutrition of the free living nematode, Chiloplacus lentos were studied

by Roy (1973a, 1973b) who also described intrauterine egg development in this

species. John and Willett (1978) developed a method for age synchrony of

Panagrellus redivivus.

Reproduction in nematodes may be accomplished by amphimixis or

parthenogenesis. A few species are also known to be hermaphrodites. Sexual reproduction between males and females of nematodes was first observed by Greet

(1964) in a free living species of nematode. Ponagrolaimus rigidus and later on by

Jones (1966) in Pe/odera leres. Green (1966, 1967) Green and Plumb (1970) not only demonstrated the presence of sex attractants in Helerodera rostochiensis and 11 schachtir but also gave an account of their orientation behaviour. Attraction between various strains of Panagrellus sp. was found to be dependent on concentration of the extract (Balakanich and Samoiloff 974). Croll (1970) analysed swarming in

Panagrellus redivivus and concluded that swarming is dependent on the characteristics of locomotion and physical forces in the environment. According to

Cheng and Samoiloff (1971) even the fourth stage of Panagrellus silusiae are capable of producing sex attractants and males of the same stage are equally capable of

32 responding to it, but copulation occurs only at maturity. Chin and Taylor (1969) and

Jairajpuri and Azmi (1977) gave conclusive evidence of the presence of sex

attractants in Cylindrocorpus sp. and Acrobeloides sp.

In 1964, Doncaster designed four patterns of observation chamber for studying

nematode behavior. Dropkin (1965) discussed the nematode behaviour in relation to

that of other invertebrates. Chuang (1962) and Thomas (1965) reported unequal

division of blastomeres for Rhabditis ieres and Acrobeles complexes respectively.

Thomas (1965) also observed that A. complexus larvae exhibited an increased number of pulsations of the memcorporal valve before emergence from the eggs. They also observed the development of the anterior half was arrested in one of the eggs under observation and it resulted in an embryo with a well developed posterior half and a mass of undifferentiated cells in the anterior half. The posterior half showed vigorous movements for 24 hours before death occurred. Their generation time was also observed which was found to be less than 2 days at 30°C for both the species.

Nicholas (1962) reported generation time as 7-8 days for Acrobeloides huet.schll at

30°C and Cryan et at (1963) reported generation time of 5-6 days for Panagrellus redivivus at 25°C.

Studies of the diplogastrids are mainly of their predatory behaviour. Aspects of reproduction and developmental biology was studied by Triantaphyllou and

Hirschmann (1964), Pillai and Taylor (1968) and Hechler (1970). Pillai and Taylor

(1968) studied in detail the biology of Paroigolaimella bernensis and Ficmr anchicoprophaga in laboratory culture. They reared predacious nematodes -

Mononchoides spp., Ficlor anchicoprophaga and Pullerius sp. in a dixenic culture of bacteria and amoebae as well as on the nematode AphClCnChIIS avenge. Members of these genera were observed to behave cannibalistically in the abseneC of other food or when the supply of food was scarce as older cultures. Pillai and Taylor (1968) also

33 recorded progeny production of five females of Paroigolaimella bernensis and Fictor

anchicoprophaga to average 18,000 after 10 days at 20°C.

Chin and Taylor (1970) described molting in species of Cylindrocorpus in

detail. Rogers (1962) suggested that molting in free living nematodes might be

controlled by internal secretions, whereas in parasitic forms this process is replaced

by host stimuli. Jantunen (1964) and Hechler (1967) have described exsheathment

patterns in C. briggsae and Diploscapier coronata respectively. Sommerville (1957)

reported that several animal parasitic nematodes secreted an exsheathing fluid which

attacked the sheath at a specific area near the anterior end, causing a refracting ring to

appear in the cuticle. Subsequently, the cap- like region of the cuticle, anterior to the

refracting ring, became detached and the namatodes escaped. But no such ring or head

cap was observed in molting specimens of Cylindrocorpus. Jantunen (1964) reported

that in molting of C. briggsae the cuticular lining of vagina was shed along with the

body cuticle, while, in Cylindrocorpus species, the vaginal lining is initially laid down

in the pre-adult female and is not shed during the molt.

Anya (1976) and Lee & Atkinson (1976) reviewed reproductive physiology of nematodes. Later, Grootaert (1976) and Poinar et al. (1976) studied the life cycles of

Mesodiplogaster lheritieri and Eudiplogaster aphodii. Chin (1977) gave a detailed account of embryonic and post-embryonic development of Cylindrocorpus longistoma. Seymour et al. (1978) designed a recording system to study the nematode behaviour. Duggal (1978a. 1978b & 1978c) discussed sex attraction in Panagrel/us redivivus and also observed the copulatory behaviour and initiation of copulation and its effect on oocyte production and life span of adult male.

Woombs and Laybourn Paery (1984) described the growth, reproduction and longevity in Diplogasteritus nudicapitatus, Hirschmanniella ory•rae and second stage juvenile of Mfeloidogyne incognita. Predatory nematodes are unable to detect their

34 prey and their predation depends on chance encounters (Yeates, 1969; Grootaert and

Maertens, 1976; Small and Grootaert, 1983; Bilgrami, Ahmad and Jairajpuri, 1984).

Esser (1963) speculated that dorylaim predators were attracted towards excised prey

from short distances. Bilgrami, Ahmad and Jairajpuri (1985) and Shafqat et al. (1987)

however, did not find any positive attraction in Aquatides thornei and Dorylaimus

stagnalis respectively towards excised prey, but observed aggregation of more than

one predator taking place around a prey injured by another predator. Yeates (1969)

and Wyss and Grootaert (1977) also reported similar aggregation in Diplenteron

potohikus and Labronema vulvepaprllatum.

Chitambar and Noffsinger (1989b) studied the predacious behaviour and life

history of Odontopharvnr longicaudata in water agar culture. The predatory nematodes use a variety of mechanisms to obtain prey. Similarly, prey nematodes possess certain qualities such as the thick cuticle, annulations, chemical secretions, etc., which may protect them from predators (Esser, 1963). Esser (1963) provided a list of nematodes that he found to be susceptible or resistant to predation by different mononchs and dorylaims. Grootaert, Jaques and Small (1977) observed prey selection in Builerius sp. and measured susceptibility and resistance to predation of tylenchid. aerolaimid. enoplid and dory laim id nematodes. Small and Grootaert (1983) described anti-predation adaptations in prey nematodes.

The diplogastrids have both parasitic (Poinar ei al., 1976) and predacious

(Bilgrami, 1997) members and possess far more beneficial biological control than any other predatory group (Bilgrami, 1997). Most diplogastrid species are small yet possess high fecundity with a short life cycle - approximately one week, the two desired traits of efficient nematode bio-control agent (Tahseen etal., 1990, 1994).

Lopez-Llorca and Olivares-Bernabeu (1995) gave a simple technique for obtaining Panagrellus redivivus from axenic cultures. Comparative studies of vulval

35 development have been initiated by cell lineage analysis and cell ablation experiments

in species of different families of nematodes (Sommer and Sternberg, 1994, 1995,

1996a). This approach was extended to genetic level in the newly described species

Pristionchus pacificus (Sommer et al., 1996). Pristionchus was the first member of

the suborder Diplogastrina to be analysed with respect to vulval development.

Sommer (1997) reported the comparative studies of cell lineage and pattern formation

during vulval development in the species Diplogaster maupasi, Koerneria sp.,

Diplogasteroides sp., Pseudodiplogasteroides sp., Aduncospiculum halicti and

Goodevus ulmi. All species display an identical differentiation pattern of ventral

epidermal cells as revealed by cell lineage analysis. Despite these morphological

similarities, cell ablation experiments suggest differences in the underlying pattern

formation processes. The finding that the pattern of cell competence varies among

species indicates that developmental mechanisms can change during evolution while

differentiation pattern remain invariant. A combination of cellular, genetic and

molecular tools have provided a detailed understanding of several developmental

processes in the free living nematode C. elegans. One aspect of development that is

tractable for comparative analysis is the generation of the vulva, the egg laying

structure of nematode females or hermaphrodites (Sommer and Sternberg, 1996b).

The vulva is a derivative of the ventral epidermis, which consists of 12 epidermal cells.

The nematode Pristionchus pacificus has been established as a satellite organism in evolutionary developmental biology (Eizinger et al.. 1999; Sommer,

2000). Developmental studies concerning P. pacificus have mainly focused on post- embr onic development, a detailed knowledge of the early development is, at best, fragmentary. Dolinski et al. (2001) studied the time of establishment of the germline cell P4 (late vs early), the arrangement of blastomeres in the four cell stage (linear,

36 partial linear or oblique) and whether AB and Pl initially develop at the same

(synchronous) or different (asynchronous) rates. A comprehensive study on the

similarities and differences in the early development of nematode embryos was made

by Schierenberg (2000). Furst von Leiven (2003b) provided a detailed account on

functional morphology and evolutionary origin of the three-part pharynx in

nematodes.

Recently, developmental and molecular studies of P. pacificus ha ve been

complemented by genomic data comprising the genetic linkage map (Srinivasan et al.,

2002). the physical map (Srinivasan et al., 2003), and whole genome sequencing,

which is approaching completion. In addition, the ecology and the specific

habitat/ecological niche of P. pacificus have been studied. Comparative studies

between P. pacificus and C. elegans have revealed differences in the regulation of developmental processes such as the development and genetic analysis of sex determination (Piresda Silva and Sommer, 2004) vulva development (Sommer and

Sternberg, 1994, 1996b: Sigrist and Sommer 1999; Sommer, 2005) and gonad formation (Rudel et al., 2005: Sommer. 2005). Furst von Lieven (2005) compared the timing of developmental events between Caenorhabditis elegans and Pristionchus paci/icus and concluded that diplogastrids have only three postembryonic preadult stages compared to the four usually present in nematodes. Sommer et al. (2005) studied embryonic, gonad, vulva and male tail development in detail and compared in nematodes of five different families and also provided a detailed picture of evolutionary changes in development.

Herrmann et al. (2006) sho"ed that some Pristionchus sp. are often associated with scarab beetles and the Colorado potato beetle. To interpret these ecological patterns, a phylogenetic framework of the genus Pristionchus was recently presented

(Mayer et al., 2007). Vangestel, llouthoofd. Bert and Borgonic (2008) provided the

37 details about the early embryonic development of the satellite organism Pristionchus

pacificus and also observed its differences and similarities with C. elegans. Bar-Eyal

et al. (2008) suggested Koerneria sudhausi as a biocontrol agent for root knot

nematodes as it reduced the galling index of tomato plants caused by Meloidogyne

jativanica in soil. Schulze and Schierenberg (2011) provided a detailed discussion on

evolution of embryonic development in nematodes by using 4-D microscopy and 3-D

modeling of developing embryos.

In India, the work on biology of nematodes was virtually untouched except for

the work done by Jairajpuri et al. (1976, 1977) and Azmi and Jairajpuri (1976, 1977).

Jairajpuri and Azmi (1977) gave a good account of the reproductive behaviour of

Acrobeloides sp. Ahmad and Jairajpuri (1977) studied the developmental biology of

Chiloplacus symmetricus. Ahmad and Jairajpuri (1980) studied sex attraction and copulation in Chiloplacus symmetricus in detail.

Shafqat. Bilgrami and Jairajpuri (1987) observed prey selection in

Dorvlaimus stagnalis Dujardin, 1845 and suggested that thick cuticle of Hoplolaimus indicus Sher. 1963 and chemical secretions of Helicotylenchus indicus Siddiqi, 1963 may represent anti-predatory devices. Bilgrami and Jairajpuri (1988) observed the pre and post-feeding aggregation of Mononchoides longicauclatws and M. fortidens at feeding sites and attributed this phenomenon to lingering prey attractants in that area.

The predatory mononchs, however, did not show such a response towards their prey

(Bilgrami, Ahmad and Jairajpuri, 1984). Tahseen and Jairajpuri (1988) provided the detailed description and developmental biology of Teratorhabditis andrassyi.

Bilgrami and Jairajpuri (1988) studied in detail the attraction of Mononchoides longicaudatus and At. fortidens towards prey and factors influencing attraction on agar plates. These predators were attracted towards excised and non-excised (live) prey in response to prey secretions, attractants. Predators also responded positively

38 towards agar containing unidentified bacteria. M. forlidens showed preference for the

prey nematodes which were tested either alive or by cutting into pieces (excised).

Both species of predators were also attracted towards their own individuals. Different

factors viz.. prey number, temperature, incubation period of prey, starvation of

predators and distance of predators from prey affected attraction of the predators.

Resistance of prey nematodes to predation, strike rate of the predators, Mononchoides

longicaudatus and M. fortidens and their ability to wound prey and feed upon them

was determined on agar plates by Bilgrami and Jairajpuri (1989a, 1989b). Tahseen el

al. (1990) provided the details of the life cycle of Mononchoides fortidens with

emphasis on gonad development. Tahseen et al. (1991a, 1991b) observed the

developmental biology of Cephalobus parvus and also studied the embryonic and

post-embryonic development of Diploscapter orientalis. Siddiqi et al. (2004) studied

biology of Mononchoides gaugleri in detail. Bilgrami et al. (2005) observed prey

preferences and feeding behaviour of Mononchoides gaugleri. Tahsecn and Nisa

(2006) studied embryology and development of gonads in Oscheius shamind. Despite

recent researches on the model organisms- Caenorhabditis elegans, Pristionchus

pacificus our knowledge of their diversity, basic nematode biology and ecology

remains meager.

Nematodes represent the most numerous multicellular organism and are next

to insects in species diversity. Understanding their diversity and functional role in the subterranean ecosystem requires knowledge on their biology. The diplogastrid nematodes form an interesting and unique group within the order Rhabditida of the phylum Nematoda. These nematodes share many features common with the order

Tylenchida while retaining their basic rhabditid features. Like Caenorhabditis elegans. several diplogastrid species are now used as models in experimental biology like developmental biology. Pristionchus pacificus is one such species in the suborder

39 Diplogastrina that is comparable to C. elegans. Occurring in decayed and decaying

organic matter diplogastrids are very frequently associated with insects which act as

phoretic hosts transporting them from one place to another. While most insect

associations are external, there are some species that occur within the insect body.

Pathogenesis to insects has not been reported so far. Feeding habit in this group is

very diverse ranging from free-living fungal and bacterial feeders to predators. They

may also be insect associates. The predatory nematodes like Mononchoides have been

used as successful biocontrol agents against plant parasitic nematodes (Bilgrami and

Jairajpuri, 1989). There could be many more such species in this one of the least

studied groups of nematodes in such a topographically diverse country. Studies on the

taxonomy and biology may result in finding more suitable models of experimental

biology or biocontrol agent.

In India the work on information on the taxonomy and biology of

diplogastrids is very restricted and meagre. Not many workers have concentrated on

this important but difficult group of nematodes on the global platform also. To

understand the role of diplogastrids in biodiversity, a sound knowledge and

understanding of its taxonomy together with details of its biology and behaviour is essential. It is. therefore. planned to investigate the taxonomy and biology of some species of diplogastrids occurring in India. In the present work we have followed the classification of Andra'ssv (1984) with some modifications. In all, twenty-three species belonging to fourteen different genera falling under seven subfamilies under five different families under two superfamilies of the suborder Diplogastrina have been described. Of these. one genus and fourteen species which are new to science and nine already known species have been described and illustrated in detail. Five of the total fourteen genera and four of nine already known species have been reported for the first time from India. The embryonic and post-embryonic development of

40 Myctolaimus oreodoxus sp. n. has also been studied in detail. The process of fertilization, egg-laying, cleavage pattern, activities of first stage juveniles within the egg, hatching, moulting and copulation were also observed. The terminology used in the text to describe the parts of stoma is of De Ley et al. (1995) and of other structural details of Ftirst von Lieven and Sudhaus (2000).

41

Soil sampling: Samples for diplogastrid nematodes were collected from farmyard

manure, chicken manure, leaf-litter, compost, decayed and decaying plant parts and

soil samples rich in organic debris from different parts of the country, especially from

the northern and north-eastern regions (Jammu and Kashmir, Uttar Pradesh,

Uttarakhand and Manipur). The samples were collected in different seasons from

different types of ecosystems like, hills, mountains, lakes, rivers, canals, ditches,

ponds, tree-holes, forests, cultivated soil etc. so as to collect all the nematodes

prevalent in different seasons and in different ecosystems. "[he samples were collected

from the top 10 — 15 cm and kept in airtight polythene bags. All relevant information

such as host, locality, and date of collection were noted and the samples were brought

to the research laboratory for further processing and studies.

Processing of samples: The samples were processed by modified Cobb's (1918)

sieving and decantation and modified Baermann's funnel techniques. Around 500 cc

of sample was placed in a bucket and mixed with water thoroughly. The debris and pebbles, if present, were removed and soil crumbs (in case of soil samples) were broken manually. The bucket was then filled with water and the suspension was stirred thoroughly to make it homogeneous. It was left undisturbed for about half a minute so as to allow the heavy soil particles to settle down to the bottom of the bucket. The suspension was then passed into another bucket through a coarse sieve (2 mm pore size), which retained debris, roots and leaves. The suspension in the second bucket was again stirred thoroughly and left for another half a minute and then poured through a 300-mesh sieve (pore size 53 pm). The catch on the sieve containing nematodes and very fine soil particles/organic matter was collected in a beaker. The process was repeated two to three times for good recovery of nematodes.

42 Isolation of nematodes: The residue collected in the beaker was poured on a small

coarse sieve lined with tissue paper and placed in a Baermann's funnel fitted with

stoppered rubber tubing. Water was slowly poured into the funnel until it touched the

bottom of the sieve. Care was taken to avoid trapping air bubbles at the bottom of the

sieve. The nematodes migrated from the sieve into the clear water of the funnel and

settled at bottom. After 24 hours a small amount of water containing the nematodes

was drained from the funnel into a cavity block. The nematodes thus isolated were

fixed and processed for mounting on slides.

Killing and fixation: The nematodes collected were le(i undisturbed for some time so

as to allow them to settle to the bottom of the cavity blocks. Excess water was then

removed with a fine dropper and hot FG (8 ml of 40% commercial formaldehyde + 2

ml of glycerol + 90 ml of distilled water) was poured into the nematode suspension.

This simultaneously killed and fixed the nematodes.

Dehydration, mounting and sealing: After 24 hours of fixation the nematodes were

transferred into a mixture of glycerine-alcohol (5 parts glycerine + 95 parts 30%

alcohol) in a small cavity block, which was then kept in a desiccator containing

anhydrous calcium chloride. Occasionally the fixative was removed as much as

possible with a dropper and glycerine-alcohol was added. In this way the entire

suspension could be processed to glycerine. In 3-4 weeks the nematodes were

dehydrated and ready to be mounted. A small drop of anhydrous glycerine was placed on a clean glass slide and the nematodes were transferred from the cavity block into

this drop. Either a ring of wax was made or the pieces of wax were kept around the drop and a circular glass cover slip was gently placed on the ring or wax pieces and the slide was kept on a hot plate to allow the wax to melt and seal the slide.

43 Preparation of the culture media: The nematodes were cultured in 1% (1 g of agar

in 100 ml of water) and 1.5% (1.5 g of agar in 100 ml of water) agar medium in 5 cm

and 10 cm diam. petridishes. The nematodes were inoculated into the agar with a

picking needle made up of a bamboo splinter or a horse hair. Approximately 5 mg of

milk powder (Lactogen) was sprinkled over the surface of agar to enhance the growth

of the bacteria upon which the nematodes fed. No attempt was made to sterilize the

medium or the nematodes.

Handling of Nematodes: The nematodes were recovered from the culture by the

modified Baermann's funnel technique whenever required. The agar containing the

nematodes was cut into small pieces and placed on a sieve lined with moist tissue

paper. After 24 hours majority of the nematodes settled at the bottom which was

extracted in a cavity block. Picking was done with the help of a picking-needle.

Observation chambers: Freshly laid eggs were placed on an agar strip in cavity

slides and observed under microscope for embryogenesis and cleavage pattern.

Embryonic development was also studied in hanging drops of water.

Staining: To stud-, the post-embryogenesis (gonad development) live and fixed juveniles of all stages were stained in 1.5% lacto-aceto-orcein (1.5 g orcein + 33 ml lactic acid + 33 ml acetic acid + 34 ml distilled water). Over-stained nematodes were transferred to 45% lactic acid for about 4-5 minutes for destaining. Staining enables proper distinction of different kinds of nuclei. The cytoplasm does not take up the stain. The stained specimens were mounted in a drop of dilute stain for observation at a higher magnification.

44

Measurements and drawing: All measurements were made on specimens mounted

in dehydrated glycerine with an ocular micrometer. De Man's (1884) formula for.

denoting dimensions of nematode was used. Morphological observations and

drawings were made on a Nikon 80i DIC microscope and photographs were taken by

a ProgRes C3 camera mounted on an Olympus BX 50 DIC microscope.

Abbreviations used in the text

L = Total body length

a = Body length / greatest body diameter

b = Body length / distance from anterior end to the oesophago-intestinal junction

c = Body length / tail length

c" _ "Tail length / anal body diameter

V = Distance of vulva from anterior end X 100 / body length

T = Total length of testes X 100 / body length

ABD = Anal body diameter

VBD = Vulval body diameter

GP = Genital papilla/papillae

Diam. = Diameter/diameters

Anterior pharynx = Anterior end to base of median bulb

45 i ORDER RHABDITIDA Chitwood, 1933

Diagnosis: Cuticle usually annulated, rarely ornamented with longitudinal striae or

punctuations. Labial region mostly continuous. Lips three or six, rarely four in

number. Amphids on the lateral lips, pore-like. rarely circular or slit-like and rarely

post-labial. Stoma prismatic, longer than wide. composed of three basic elements vi=.,

cheilostom, gymnostom and stegostom. Stegostom with denticles or well developed

teeth or fine warts. Pharynx with either median or terminal valvular bulb. Excretory

system consisting of a double collecting canal connected to a common duct. Intestine

with wide lumen. Three rectal glands generally present. Female reproductive system

amphidelphic or mono-prodelphic, in latter case generally with post-uterine sac.

Ovaries reflexed. Ovo- or viviparous. Males with paired genital papillae. Bursa may

be present or absent, if present having paired rod-like papillae or ribs. Spicules occasionally fused distally. Phasmids distinct. Tail often distinct in each sex, without caudal glands or spinneret.

Type suborder: Rhabditina Chitw•ood, 1933

Other suborders: Cephalobina Andrassy. 1974

Diplogastrina Micoletzky, 1922

Teratocephalina Andrassy, 1974

SUBORDER DIPLOGASTRINA Micoletzky, 1922

Diagnosis: Lips mostly six, less separate. Amphids mostly distinct, particularly in males, oval, in the anterior stomal region or in the posterior part of the stoma and sometimes, still beyond it. Stoma divisible into cheilostom, gymnostom and stegostom: tubular or wide barrel-shaped; bilaterally symmetrical or asymmetrical.

Stegostom always with asymmetrical swellings; the dorsal usually stronger and

46 differently structured than the subventrals and often with large, movable claw-like

tooth. Pharynx divisible into two parts. The anterior pharynx comprising of the

muscular procorpus and strong median bulb housing the crescent valve plates and the

posterior pharynx of isthmus and the glandular terminal bulb. Female gonads mostly

paired. rarely unpaired. Ovaries reflexed. Testis one, reflexed. Spicules free, rarely

fused. Gubernaculums present. Bursa present or rudimentary or completely absent.

Caudal papillae papillose or setae-like. Tail in both sexes long filiform. Phasmids

distinct.

Type superfamily: Diplogastroidea Micoletzkty, 1922

Other superfamilies: C%Iindrocorporoidea Goodey, 1939

Odontopharyngoidea (Micoletzky, 1922) Andrassy, 1984.

SUPERFAMILY DIPLOGASTROIDEA Micoletzky, 1922

Diagnosis: Lips six. Labial sensilla papillae-like and males with additional four fine

cephalic setae. Amphidial openings either small, indistinct or large in the posterior

region of stoma. Cheilostomal walls mostly cuticularized. Gymnostomal walls cuticularized. Stegostomal walls asymmetrical, dorsal vall with armed tooth. Anterior

pharynx always with valvate median bulb and posterior pharynx with distinguishable terminal bulb. Female gonads mostly paired, rarely unpaired. Bursa present or rudimentary or absent.

Type family: Diplogastridae Micoletzky. 1922

Other families: Diplogasteroididae Filipjev & Schuurmans Stekhoven, 1941

Heteropleuronematidae (Andrassy, 1970) Andrassy, 1984

Neodiplogastridae (Paramonov, 1952) Andrassy, 1984

Pseudodiplogasteroididae Korner. 1954

47 Tylopharyngidae Filipjev, 1934.

Family Diplogasteroididae Filipjev & Schuurmans Stekhoven, 1941

Diagnosis: Amphidial openings usually large, conspicuous in the upper part of stoma

in both sexes. Stoma oblong, more or less tubular, always longer than wide.

Cheilostom small with weakly or not cuticularized walls. Gymnostomal walls usually

cuticularized. Stegostomal walls with small inconspicuous denticles. Anterior pharynx

muscular with strong median bulb, posterior pharynx with glandular terminal bulb.

Ovaries generally paired. Bursa if present rudimentary or absent.

Type subfamily: Diplogasteroidinae Filipjev & Schuurmans Stekhoven, 1941

Other subfamily: Rhabditolaiminae Andrassy, 1984.

Subfamily Diplogasteroidinae Filipjev & Schuurmans Stekhoven, 1941

Diagnosis: Stoma cylindrical, tubular or wide barrel-shaped. Amphidial openings in

both sexes large, conspicuous, in the middle of stoma. Female reproductive system

paired or unpaired. Bursa completely reduced or absent.

Type genus: Diplogasteroicles De Man, 1912

Other genera: Goffartia Hirschmann. 1952

Paramonovnema Andrassy, 1968

Genus Goffartia Hirschmann, 1952

Diagnosis: Body 0.6 — 1.3 mm long. Lips six, fused, lip region low. Papillae small, papillose. Stoma barrel-shaped. Cheilostomal walls weakly cuticularized, gymnostornal walls short. cuticularized. Dorsal stegostomal tooth small. Amphidial openings large, oval, in the mid stomal region. Posterior pharynx longer than the

48 anterior pharynx. Female reproductive system amphidelphic. Spicules small, arcuate.

Bursa rudimentary or absent. Tail long filiform in both sexes.

Type species: G. heteroceri Hirschmann, 1952

Goffarlia baldus sp. n. (Fig. 1, 2)

Measurements: In Table 1.

Descriptions

Females: Body small, always less than 1 mm, slender, almost straight upon fixation.

Cuticle thin, transverse striations not visible. Subcuticle finely striated. Lip region

round, smooth, continuous with body contour. Lips fused, labial papillae not

discernable. Amphidial apertures large transverse slits with an indentation in the

middle, almost as wide as corresponding body, about one lip diam. from anterior end.

Amphidial pouches (fovea) large, wine glass shaped, almost as wide as lip diam., amphidial canal short, fusus slightly swollen. Lateral fields and lateral lines obscure.

Stoma narrow, tubular. Cheilorhabdions weakly sclerotised. Gymnostom barrel- shaped about as long as cheilostom. Stegostom short, without tooth or denticles.

Procorpus muscular. Isthmus broad, long, gradually widening in the posterior third to form the glandular basal bulb. Ratio of anterior pharynx (from anterior end to base of median bulb) to posterior pharynx 1:1.6-1.8. Nerve ring in the middle or posterior half of isthmus, at 55 — 60 % of pharyngeal length. Excretory pore obscure, hemizonid not visible. Intestinal cells small.

Reproductive system amphidelphic, anterior branch on right and posterior on left side of intestine. Ovaries reversed, oocytes in one or more rows in the germinal zone. Oviduct long, narrow, distally expanded to form the spermatheca, usually set off from the uterus by a constriction. Uterus divisible into glandular and muscular parts.

49 Vagina muscular, with two pairs of unicellular glands opening at the junction with

uterus, Vulval opening small, oval, transverse. Rectum 1.2 anal body diam. long.

Phasmids large, prominent, 3.0 - 3.5 anal body diam. posterior to anus. Tail very long,

whip-like, tapering to a pointed tip, 1.1-1.6 times vulva-anus distance long.

Mates: Similar to females in general morphology. but smaller in size and with

posterior region strongly curved. Lip region like in females, cephalic papillae/setae

not visible. Spicules narrow, ventrally arcuate. Gubemaculum broad, keel-shaped,

with a distal sleeve. Genital papillae nine pairs; two pairs pre-cloacal, seven pairs

post-cloacal. Genital papillae formula: (vl, v2)/ v3d, v4, ad, phasmids, v5 (v6, v7), pd. Papilla v5 of the v5-7 group placed anteriorly at level of ad. Tail with a short conoid part and a long filiform part.

Type habitat and locality: Farmyard manure collected from District Baramulla,

Kashmir, J&K,

Type specimens

Holotype: Female on slide Goffartia baldus sp. n./8 deposited in the nematode collection of the Department of Zoology, Aligarh Muslim University, Aligarh.

Paral pes: Twelve females and seven males on slides Gofjhrlra baldus sp. n./I-7 deposited in the nematode collection of Section of Nematology, Department of

Zoology, Aligarh Muslim University, Aligarh.

Diagnosis and relationship

Goffarlia baldus sp. n. is characterized by a small slender body, large amphidial apertures and wine glass shaped fovea; a smooth round lip region without discernable labial papillae; a tubular stoma with almost equal cheilo- and gymnostoms; pharynx with a short corpus and a long post-corpus; two pairs of unicellular glands at base of vagina, large prominent phasmids; a broad keel-shaped gubernaculum with distal sleeve and nine pairs of genital papillae.

50 The new species is easily distinguished from all other species of the genus by the broad keel-shaped gubernaculum, unicellular glands associated with the vagina and the smooth, round lip region without discernable labial papillae. Further differences with G. heteroceri Hirschmann, 1952, G. praepilata Shoshin. 1989, G. africana (Micoletzky, 1915) Hirschmann, 1952 and G. frlicaudata (Andrassy, 1968)

Sudhaus & Furst von Lieven, 2003 is in the proportion of the anterior and posterior pharynx (1:1.6-1.8 vs approximately 1: 1 in others; as interpreted from figures).

Etymology: The species name is after the smooth, rounded lip region.

51 Table 1: Measurements (in µm) of Goffartia baldus sp. n. Mean and S. D. given in parenthesis.

Characters Holotype Paratype females (n = 12) Paratype males (n = 7)

L 625 573-647(615±21) 450-574(511±34.5)

a 39.5 37.5 - 44.7 (40.2 t 2.2) 39.7-48.3 (43.5 ± 2.5) b 5.5 4.9-5.7 (5.4 ± 0.2) 4.2 - 5.2 (4.7 ± 0.4)

c 2.94 2.7 - 3.5 (3 ± 0.2) 2.9-3.7 (3.3 ± 0.2)

c' 23.89 18.3-26.3 (22.5 ± 2.1) 12.4-19.7 (15.5±2.2) V 41.46 40-45(42± 1.5) Maximum body width 16 14-17(15.5 ± 1.0) 10.9-12.9(11.8 ± 0.6)

Lip width 5 5-6(5±0.3) 5 Length of stoma 5 5 - 7 (5.5 ± 0.7) 5 -6 (5.2 ± 0.4) Anterior pharynx 45 43.5 - 48.5 (46.0 ± 1.5) 42 - 46 (43.5 ± 1.5) Posterior pharynx 69 62 - 76 (68.5 ± 3.5) 54 -71 (66 ± 5) Pharynx 114 107-124(114.5±4.5) 96-117(109±6) Anterior/posterior pharynx 0.65 0.62 - 0.74 (0.67 f 0.03) 0.6 -0.8 (0.7 f 0.04) Nerve ring from anterior end 69 62.5 - 76.5 (69.5 f 3.5) 59 -73 (65 t 4) Median bulb (width) 6 6-7 (6.5 ± 0.5) 6 Anterior gonad 60 55 - 73 (64.5 ± 5) Posterior gonad 74 45 - 83 (64 ± 10) Vulval body diameter (VBD) 16 14- 17 (15.5 ± 1) Vulva-anus distance 154 146-177 (154 ± 8) Testis (length) 129-203 (162 ±21) Spicules length (along axis) 14.8 - 15.8 (15 ± 0.4) Gubernaculum length 7

Rectum 10 10-12(11± 1.0) 14-18(16± 1.5)

Phasmids from anus/cloaca 26 26 - 32 (28.5 ± 2) 18 - 22 (19.5 ± 1.5) Tail ' 213 163-235(204±18) 123-195(158±20) Anal body diameter (ABD) 9 9- 10 (9.1 ± 0.4) 10- 11 (10.2 ± 0.4)

52

i'D E1~ F

i

K \ \)k; \

L. i----s 10 Pill

K~ 20µm

Fig. 1. Goffarlia baldws sp. n. A. Entire temale. 13. Entire male. C. Anterior end, D. Phary nx, F. Male anterior end showing amphids laterally, F. Female anterior end shoo\%ing aniphids laterally. G. Anterior region (dorsoventral). II. Female Reproductive System (entire). I. Inicellular Glands, J. Female posterior region. K. Male posterior region. L. Spicules. M. Gubernaculrnn.

;3 A 41r C 7

~

E

Fig. 2. (io//Irtia haldus sp. n. A. Pharyngeal region. B. D. Anterior region showing amphids (lateral). C. Anterior region showing stoma. E. Female Reproductive system (entire). F. Female Reproductive ssstem showing eke.. G. Vulval region showing unicellular glands. II. Testis. I. J. Male posterior region showing spicules and gubernaculum. (Scale bars - B. C. D. J = 10 µm. A. F-I 20 µm). Subfamily Rhabditolaiminac Andrassy, 1984

Diagnosis: Stoma cylindrical, rhabditid-like. Amphids small, inconspicuous at the

base of lips or in mid-stomal region. Female reproductive system paired or unpaired.

Bursa present or rudimentary.

Type genus: Rhabditolaimus Fuchs, 1915

Other genera: Rhabditidoides Rahm, 1928

Rhabdontolaimus (Fuchs, 1931) Filipjev & Schuurmans Stekhoven, 1941

Fuchsnema Andrassy, 1984.

Genus Rhabditidoides Rahm, 1928

Diagnosis: Body length ranges from 0.6-2.3 mm. Lip-region low, papillac medium-

sized. Stoma small, cylindrical, 5-6 times as long as wide. Cheilostomal walls

pointed, comma-like. Gymnostom tubular. stegostom with dorsal flap-like process.

Female reproductive system amphidelphic. Genital papillae 9 pairs; 3 pre and 6

postcloacal. Spicules sickle-shaped. bursa absent.

Rhabditidoides papillatus Mahamood and Ahmad, 2009 (Fig. 3, 4)

Measurements: In Table 2.

Descriptions

Females: Body medium sized, tapering towards both extremities. Cuticle with fine transverse and longitudinal striations. Punctations arranged both in transverse and longitudinal rows. Lip region continuous with body contour. Lips fused with weekly developed labial papillae. Amphidial apertures oval, at junction of cheilostom and gymnostom. Stoma tubular, longer than wide. ventro-dorsally curved in posterior region. Cheilostom with weakly cuticularized cheilorhabdions. Gymnostom anisotopic and anisomorphic. Stegostom anisomorphic, dorsal wall with a large,

55 ventrally directed tooth. Posterio-ventral aspect strongly cuticularized. Anterio-dorsal

aspect with an uneven surface. Subventrals with a cuticularized ridge each. Procorpus

muscular, median bulb oval-shaped with large valve plates. Isthmus short expanding

posteriorly to an ellipsoidal basal bulb. Anterior pharynx 68 — 72 % of pharyngeal

length. Nerve ring encircling isthmus in mid or in anterior half. Hemizonid located

posterior to the nerve ring, 76 - 88 % of total pharyngeal length. Excretory pore

slightly posterior to hemizonid. Dierids at the level of basal bulb. Three additional

pairs of post-deirids present- first pair posterior to cardia, second pair far posterior to

vulva and third pair slightly anterior to vulva. Cardia small conoid. Intestinal cells

large and granular. Intestinal lumen generally wide.

Reproductive system amphidelphic, anterior branch on right and posterior

branch on left side of intestine. Ovaries reversed, oocytes arranged in more than one

row in germinal zone. Oviduct narrow. Spermatheca irregular in shape, larger and

wider than uterus and clearly set off from the uterus. Uterus divisible into glandular

and muscular parts. Vagina muscular, tubular, one-third of body diam. long, and thick

walled with circular vulval opening. Rectum 1.3 - 2.0 anal body diam. long. Vulva-

anus distance about 7.0 to 9.2 times vulval body diams long. Phasmids I - 2 anal body

diam. posterior to anus. Tail long filiform. 0.9 - 1.1 times vulva-anus distance.

Hales: Body curved in posterior region. Anterior region similar to that of females

except for four small cephalic setae. Testis ventral, flexure on right side. Spicules

strongly developed, arcuate with bibbed capitulum. Gubernaculum slender, distal end

with a sleeve and proximal end bifid and anteriorly directed, 48 - 62 % of spicules

length. Genital papillae nine pairs; three pairs precloacal, six pairs postcloacal.

Genital papillae formula: vi, v2, v3d / v4, (ad, v5, v6, phasmids), pal v7. Genital

papillae vI, v2 within the range of spicules when retracted. GP v3d at the level of cloaca. GP v4 far posterior to cloaca. Genital papillae ad slightly anterior to or at the

56 level of GP v5. Phasmids almost at the level of GP v6 or slightly posterior to v6. GP

v7 far posterior to GP v6 at the junction of conoid and filamentous parts of tail. GP pd

slightly anterior to v7. Tail in two parts; a short conoid part and a long filamentous

part.

Habitat and locality: Floating island Khornapaat, Loktak Lake, Manipur.

i'oucher specimens

Ten females and ten males on slide Rhabditidoides papillatu.s/l -6; deposited in the nematode collection of Department of Zoology, Aligarh Muslim University,

Aligarh.

Remarks

Measurements and description of our specimens conform well with that of the original specimens described by Mahamood and Ahmad (2009). However, differences were found in length of stoma (13 - 15 pm vs 16 - 18 pm), slightly shorter isthmus and in position of v 1 (posterior to the spicules head vs at the level of spicules head), position of v2 (almost at the level of upper cloacal lip vs always anterior to upper cloacal lip), position of ad (at the level of GP v5 or slightly anterior to it vs always anterior to v5).

57 Table 2: Measurements (in µm) of Rhabdilidordes papillatus. Mean and S.D. given in parenthesis.

Characters I Females (n = 10) Males (n = 10) L 586 - 878 (762.5 t 76.5) 560 - 783 (632.9 t 63.2) a J 24.7-31.6(27.9±2.5) 22.6-30.1 (25.4*2.5) b 5.5-7.5(6.3±0.x) 4.2-6.8(5.2±0.7) c 3.4-3.8 (3.5 ± 0.1) 3.6-6.0 (4.4 ± 0.7) c' 14.1-18.9(17.3± 1.3) 6.0-9.7(7.7±0.9) V 40.1-44.2(41.6±1.4) T 38.95-54.9 (47.9*4.3) Maximum body width 23 - 35 (27.6 ± 4.1) 21 - 27 (25.0 ± 2.1) Lip width 6-7(6.3 ± 0.5) 5 -7 (5.8 ± 0.6) Length of stoma 13- 16(14.9±0.8) 13- 15 (14.2 ± 0.8) Pharynx 104-138(121.6±10.0) 105-138(122.0±10.4) Excretory pore from anterior end 90-113(104.2 ± 8.1) 90-112(101.0 t 5.95) Hemizonid from anterior end 88 - 109 (99.8 ± 8.0) 86 - 108 (96.7 ± 6.1) Nerve ring from anterior end 83 - 104 (94.6 ± 7.2) 81 - 104 (92.4 ± 6.2) Median bulb (length) 17-23(19.9± 1.6) 17-21(18.3±1.4) Medianbulb(width) 13-17(15.0±1.4) 11-15(13.8±1.3) Pharynx base to gonad 47-99(74.0± 15.0) 17-71(48.7± 15.2) Anterior gonad 73 - 160 (121.4 ± 28.1) Posterior gonad 55 - 148 (98.4 ± 23.5) VBD 23 - 36 (29.0 ± 4.3) Vulva 251 - 364 (317.0 ± 28.9) Vulva - anus distance 165 - 278 (229.5 ± 31.2) Rectum 15-22(18.1±1.8) 15-19(16.9±1.3) i Tail 1 170-245(216.0±20.2) 120-175(147.0±17.6) ABD 11-14(12.5±0.8) 17-22(19.2±1.6) Phasmids from anus/cloaca 13-24(17 ± 3.4) 17-23 (19.7 ± 1.8) Testis 250-355(301.9± 31.7) Spicules along axis 26 - 32 (29.8 ± 1.9) Spicules along cord 23 - 31 (26 ± 2.24) Gubernaculum 15 - 17 (16.2 ± 0.8)

58 F

m

-C

Fig. 3. Rliubditidoides pupillutus. A. Entire female, B. Entire male, C. Pharyngeal region. D. Anterior region showing amphids dorsoventrally, E. Femal'. Reproductive tract (Anterior), F. Female posterior region, G. Male posterior region.

59 0

1: •!, V \ • ' - . ft

at 43

{ '3;; . 1t2' Ar 4 ).r' .r

:~ !ftitr k t

F

I

G E 1

4. R1,Lth 1ricl~rdc papa/latu.s. A. Anterior region sho lug stoma, B. Pharyngeal region. C. Female Reproductive system (anterior branch), D. Female Reproductive system shoving eggs. E. Vulval region, F. Male posterior region showing spicules and cgubernaculum, G. Male posterior region showing genital papillae (Scale bars 20µm). Family Diplogastridae Micoletzky, 1922

Diagnosis: Amphidial openings small, anteriorly situated. Cheilostomal walls

cuticularized, often divided into rods. Gymnostom spacious, rarely tubular with

cuticularized walls. Stegostomal walls with prominent conspicuous teeth. Dorsal tooth

immovable, always bigger than the subventral teeth. Pharynx' muscular, with distinct

median bulb and terminal bulb. Female gonads mostly paired. Bursa nearly or always

rudimentary.

Type subfamily: Diplogastrinae Micoletzky, 1922

Other subfamilies: Demaniel line Paramonov, 1951

Paroigolaimellinae Andrassy, 1976

Subfamily Diplogastrinae Micoletzky, 1922

Diagonosis: Stoma mostly spacious, cheilostom simple, uniform, not or rarely

divided. Gymnostom with unequal walls, the ventral longer than the dorsal.

Stegostomal walls with prominent dorsal tooth and two small comma-like subventral

teeth. Median bulb spherical. Ovaries paired, seldom unpaired. Bursa mostly absent.

Type genus: Diplogasrer Schultze in Carus, 1857

Other genera; Acroslichus Rahm, 1928

Butlerius Goodey, 1928

Diplogastre/lus Paramonov, 1952

Pseudacrostichus gen. n.

Genus Acrosticlus Rahm, 1928

Diagnosis: Body 0.5-1.9 mm long. Cuticle distinctly longitudinally striated Labial papillae small or setae-like. Stoma elongate, about twice as long as wide. Cheilostom with six adradial flaps, separated into six adradial plates. Gymnostom anisotopic,

61 stegostom anisomorphic. The subventral walls longer than the dorsal. Dorsal wall

with a large, thom-like or dagger-like tooth and a small plate at base. Female

reproductive system amphidclphic. Vulva pre-equatorial. Vagina linked to a large

kidney-shaped chamber situated opposite to vulva (absent in A. dendrophilus).

Spicules Gee, gubernaculum massive. 9-10 pairs of genital papillae, vi and v2 closely

associated, bursa absent. Tail very long.

Type species: A. loledoi Rahm, 1928.

Acrostichus nudicapilatus Steiner, 1914 (Fig. 5, 6)

Measurements: In Table 3.

Descriptions

Females: Bo dy small, tapering towards both extremities. Cuticle with prominent

longitudinal ridges and fine transverse striations. Lip region continuous with body

contour. Lips six, not distinctly differentiated, bearing a small papilla each. Amphidial

apertures circular, at level of dorsal tooth. Cheilostom cuticu!arized, narrowing

anteriorly; gymnoslom anisotopic, dorsal wall shorter than the subventrals. Stegostom

anisotopic and anisomorphic. Dorsal wall with an anteriorly directed tooth, each

subventral wall with a small tooth. Procorpus muscular, median bulb ovoid, 14-20 µm

long. Anterior pharynx 59 — 64 ?' of total pharyngeal length. Isthmus long, basal bulb

ovoid. Nerve ring encircling middle of isthmus, located at 72 — 77 % of pharyngeal

length. Hemizonid posterior to nerve ring at about 76 — 93 % of pharynx length.

Excretory pore slightly posterior to hemizonid at 78 — 95 % of total pharyngeal length. Cardia small. Intestinal lumen wide.

Reproductive system amphidelphic, anterior gonad on right side and posterior on left side of intestine. Ovaries reversed, may or may not cross each other at vulva)

62 region. Oocytes arranged in a single row except at proliferation zone. Spermatheca

not well set off from uterus containing sperms; uterus divided into two parts, a short

muscular part and a long glandular part. Vagina muscular, one-forth of vulva! body

diam. A pouch-like structure present opposite to vulva and opens into reproductive

tract at uterus-vagina junction. Vulval opening transverse, oval. Vulva-anus distance

about 5.2-5.96 vulva! body diams long. Phasmidial openings distinct, 1.2 - 1.7 anal

body diam. posterior to anus. Rectum 1.2 - 1.6 anal body diam. long, Tail, 6liform,

4.7-8.3 vulva-antis distance long.

Males: Body slightly smaller than females. Anterior region similar to that of females except for four cephalic papillae present posterior to circlet of six labial papillae.

Testis ventral, flexure on right subventral side of body. Spicules strong, arcuate, cephatated 1.6-1.8 times anal body diams tong. Gubemaculum massive, broader than spicules, 75-85 % of spicules length. Tail divided into two parts, a short, conoid part and a long filamentous part. Genital papillae nine pairs; three precloacal and six postcloacal. Genital papillae formula: (v1, v2), v3d / v4, ad, phasmids, (v5, v6, v7). pd.

Habilat and locality: Farmyard manure collected from District Baramulla, Kashmir,

J&K, India.

Voucher specimens

Ten females and ten males on slides Acrostic)rus nudicapitatus/1-9 deposited in the nematode collection of Department of Zoology, Aligarh Muslim University,

Aligarh.

Remarks

The description and morphometric measurements of our specimens concur well with that of Steiner (1914). However, a slight variation was observed in the built

63 of spicules (heavily built with blunt distal tip vs moderately built with relatively pointed distal tip).

64 Table 3: Measurements (in gm) ofAcrostichus nudi capital us. Mean and S.D. given in parenthesis.

Characters Females (n 10) Males (n = 10) L 484-615 (568.5 f 37.1) 397-500(451.2_35.8)

a 18.7-23.7(21.3+1.6) 20.7-23.9(22.0±1.1)

b 4.9-6.3(5.5 t0.3) 4.6- 5.5 (5.1 f 0.2)

c 2.9-3.8(3.310.3) 3.1-4.0 (3.5 0.3)

c 9.3- 15.8 (12.1 f1.9) 6.5-103(8.4±0.9)

V 40.2-42.8(42.1±1.2)

T 37.8-49.0 (44.9 f 3.0) Maximum body width 21-32(26.9±3.2) 18-23(20.5±1.6)

Lip width 5-6(5.6±0.5) 5-6 (5.2t 0.4)

Lipheight 2-3(2.4±0.5) 2-2(2±0)

Length of stoma 9-10(9.2x0.4) 7-8(7.2±0.4) Pharynx 98-110(102.7 + 3.8) 80-94(88 t 4.4)

Pharynx base to gonad 40-65(55.6 f8.4) 18-40(30.7x7.3) Excretory pore from anterior end 77 - 100(88.91 6.6) 70-92(79.8 + 7.0) liemizonid from anterior end 75-97(86.2 t 6.3) 68-90(78.1±7.1) Nerve ring from anterior end 71 - 80 (75.60 f 2.8) 59-77(66.2 f 4.8) Median bulb(length) 14-20(16± 1.7) 12-15(13.1±1.1) Median bulb (width) 10-14(12.1 t 1.1) 8-11(9.5±0.8)

Anterior gonad 50-102(83.1+ 13.3) Posterior gonad 50-101(76.6±135) VBD 21-31(27.3±3.2)

Vulva-anus distance 117- 177 (153 17.3) Rectum 17-21(19.3±1.2) 17-25(21.5 ± 23) Tail 140-190(173.3113.5) 110-155(131413.2)

ADD 12-17(14511.6) 14-18(15.61:1.1)

Phasmids from anusicloaca 18-21(19.510.9) 17-21(19}1.3) Testis 150-245(201.5+27.4) Spicules along axis 25 -29 (26.9 ± 1.4)

Snicule ilnnn cord 24 9± 151

65 77

Fig. 5. ?leros(icI:us iziiIicapititu.s A. Intire ternale. B. Entire male, C. Anterior region. D. Pharvngeal region. E. Female Reproductive tract (Anterior), F. Vaginal pouch. G. Female posterior region. H. Male posterior region. :► 101 D

a. L.

,,

G \\ J I. J

Fig. 6. Acrostichu.s nudicapilutus. A. Pharyngeal region, B. Anterior region showing stoma, C. Cuticle showing longitudinal ridges. I). Female Reproductive system (anterior branch). F. Vaginal pouch, F. Vuhal region showing vaginal pouch. G. Female posterior region showing amphids, H. Male posterior region, I. Male posterior reion showing genital papillae & phasmids. J. Spicules and ggubernaculum, (Scale bars 20µm). Genus Butlerius Goodey, 1929

Diagnosis: Body 0.8 — 2.0 mm long. Cuticle finely striated and mostly longitudinally

striated. Head wide, lips six in number, flat with hair-like setae. Stoma spacious with

two chambers. Anterior chamber large comprising of cheilostom and gymnostom with

cuticularized walls. Posterior chamber small comprising of stegostom. Dorsal

stegostomal wall with relatively large dorsal tooth and two small subventral teeth on

the subventral walls. Amphidial openings in males in upper part of stoma. Female

gonads paired. Spicules free, mostly slender. Gubernaculum comparatively large.

Bursa absent. Caudal papillae mostly nine pairs, setose. Tail in both sexes filiform.

Type species: B. butleri Goodey, 1929.

Butlerius n:icans Pillai & Taylor, 1968 (Fig. 7, 8)

Measurements: In Table 4

Descriptions

Females: Body medium sized, almost straight upon fixation. Body tapering slightly

anteriorly and considerably posteriorly ending in a long filiform tail. Cuticle %%ith fine

transverse and longitudinal straitions and dot-like punctations. Lateral fields with four

lines. Lip region continuous with body contour. Lips six, fused, each bearing a small

setose papilla. Amphidial opening oval, at the level of cheilostom. Stoma longer than

broad, spacious, divisible into two parts; anterior wide and posterior narrow parts.

Cheilostom wide, spacious with strongly cuticularized cheilorhabdial flaps or plates

extending beyond labial contour. Gymnostom short. anisotopic, dorsal wall shorter

than the subventrals. Stegostom anisomorphic with dorsal wall bearing a movable tooth. Right subventral %%all ith a pointed tooth and left bearing a plate with a serrated edge. Pharyngeal collar surrounds the stoma upto the stegostom. Procorpus broad, muscular, lumen zipper-like. Median bulb strong, globular, 20 - 24 tun long

and 17 - 20 tun broad. Isthmus expanding posteriorly to a glandular basal bulb.

Anterior pharynx muscular, 55 — 63 V of pharynx length. Nerve ring encircling

isthmus in anterior half, at 62 — 73 %of total pharyngeal length. I lemizonid posterior

to the nerve ring, at 67 — 81 % of the total pharyngeal length. Excretory pore

conspicuous, slightly posterior to the hemizonid, at about 79 — 97 % of the total

pharyngeal length. Cardia conoid. Intestine with wide lumen. Intestinal cells large

with clearly visible nuclei.

Reproductive system amphidelphic, reflexed with anterior branch on right side

and posterior branch on left side of intestine. Ovaries reversed. Oocytes arranged in

two or more rows in the germinal zone. Spemtatheca not offset from uterus. Uterus

with a short muscular part and a long glandular part. Vagina muscular. Vulval

opening circular. Rectum 0.9 — 1.6 anal body diam. long. Vulva-anus distance 6.7-

9.4 times vulva) body diam. long. Phasmids about 1.2 — 1.8 anal body diam. posterior

to anus. Tail filiform, 1.2 — 1.9 times vulva-anus distance long.

Males: Similar to female but smaller in body size and with four cephalic setae in

addition to six labial setae. Stoma structure and position and size of amphidial

openings similar to female. Reproductive system monorchic, testis ventral, reflexed,

flexure on right side. Spicules ventrally arcuate with bifurcated tips, 1.4 — 1.7 times

anal body diam. long. Gubernaculum slender, 42-55 % of spiculcs length, provided

with a distal sleeve. Genital papillae eight pairs; 2 precloacal and six posteloacal,

genital papillae formula: vl, v3d/v4, ad, phasmids (v5, v6, v7), yd. GP vi beyond the

spicular range when retracted. GP v2 absent. Phasmids anterior to v5. Tail long,

filamentous.

Habitat and locality: Farmyard manure collected from District Baramulla, Kashmir,

1&K, India.

69 Voucher specimens

Nine females and four males on slides Butlerius micansll-5 deposited in the nematode collection of Department of Zoology, Aligarh Muslim University, Aligarh.

Remarks

The descriptions and morphometric measurements of our specimens concur well with that of Pillai and Taylor (1968b) in general morphometrics, presence of zipper-like structure in the lumen of procorpus, in shape of spicules and number and arrangement of genital papillae. However variations were observed in c value (2.4 -

3.0 vs 3.1- 5.0), in b value in males (5.0 - 5.95 vs 6.3 - 8.7), in the position of nerve ring (in the anterior half of isthmus vs at the middle of the isthmus), in the position of excretory pore (conspicuous, slightly posterior to hemizonid vs inconspicuous, at the level of beginning of basal bulb) and in the shape of gubernaculum (with a distal sleeve vs without a distal sleeve).

70 Table 4: Measurements (in pm) ofBulle this micans. Mean and S.D. given in parenthesis.

Characters Females (n=9) Males (n = 4)

L 787-943 (869.9 t 54.5) 575-715(657.5+55.6)

a 28.5-34.4(31.8} IS) 31.95-38.3(35.3±2.5) b 5.4-7.1(6.1+0.5) 5.0-5.95 (5.5 t 0.4)

c 2.4-3.0 (2.8 t 0.2) 2.8- 3A (2.912.9)

c 15-285(21.3X4.3) 12.4-16.7(14.4±1.5)

V 36.8-40.7 (38.8 ± 1.2) T 27.8-34.2(32,132.5)

Maximum body width 23-30 (27.4 t 2,1) 15-21 (16.8±2.3) Lipwidth 13-16(14.2±1.1) 10-12(11±0.7) Length of stoma 12-14(13.210.8) 9-11(10±0.7) Pharynx 132-151(144.116.5) 115-121(118.8±2.3) Excretory pore from anterior end 114- 133 (123.3 ±6.0) 97 -100 (99 f 1.2) Hemizonid from anterior end 100-119(10646.7) 00-95(93.5+ 211)

Nerve ring from anterior end 92- 107 (96.9k 4.3) 77 -84(79.5 ± 2.7)

Median bulb (length) 20-24(21.3 f 1.4) 15-17(1610.7) Median bulb(tvidlh) 17-20(18.6± 1.4) 12-14(13.5±0.9) Pharynx base to gonad 65-105(793± 11.6) 90-120(101.3±11.4)

Anterior gonad 96-142(114+ 16.1)

Posterior gonad 85-143(117+ 19.7) VBD 3 3.1) Vulva 303-380(337.4125.1)

Vulva- anus distance 180-245(215±24.5)

Rectum 17-20(194±0.96) 16- 18(17.5 t 0.9) Tail 285-370(317.8=25.6) 200-250 (225 t 20.6) ABD 12-19(15.3 f 2.3) 14-19(15.8±1.3)

Phasmids from anus/cloaca 18-25(21.7 f 2.2) 20-22(21.3 t 0.8) Testis 160-240(212.5±313)

Spicules along axis 20-26(24 f 2.4) Spicules along cord 18 - 21 (20 f l.2) Gubemaculum II -12(11.3 f 0.4)

71 T

50 .un

Fig. 7. Butlerius micans. A. Entire female. B. Entire male. C. Pharyngeal region. D. Female Reproductive tract Anterior). E. Female posterior region. F. Male posterior region.

72 C 4 f4

: J

-.'.-.I C C r vi

t r

J K H lie.S lSri/w•iu.s ,ni zm. A. Anterior rcu,iorr int ,1rciinr, ,tcmla. It. Aiiteriur rcwofl showing serrated plate (dorsoventral). C'. Anterior region showing amphids (dorsoventral). D. Anterior pharynx. L. Median bulb, isthmus & basal bulb, F. Reflexed ovary, G. Vulval opening (dorsoventral), II. Female posterior region (lateral), I. Female posterior region showing amphids (dorsoventral), J. K. Male posterior region showing spicules & guhernaculum, (Scale bars 2O im). Genus Diplogastrellus Paramonov, 1952

Diagnosis: Cheilostom as long as gymnostom, with six flaps. Gymnostom wider than

long, anisotopic, dorsal gymnostomal wall being much shorter than subventrals.

Stegostom anisotopic and anisomorphic: dorsal wall with dorsal flap or tooth.

Subventral walls with a tooth or teeth or simply a ridge. Female reproductive system

monoprodelphic. Bursa absent.

Type species: D. gracilis (Butschli, 1876) Paramonov, 1952.

Diplogastrellus curvatus sp. n. (Fig. 9, 10)

Measurements: In Table 5.

Descriptions

Females: Body small almost straight upon fixation. Cuticle with fine transverse

striations. Amphidial apertures circular, at the base of gymnostom. Lip region

continuous with body contour, lips fused with well-developed labial papillae. Stoma

barrel-shaped, 1.8 — 2.2 times as long as lip diam. Cheilorhabdions cuticularized,

plate-like. Gymnostom anisotopic, dorsal wall shorter than the subventrals. Stegostom

anisotopic and anisomorphic. Dorsal wall with a sharp, anteriorly pointed tooth

supported by a smaller tooth at base. The subventrals with a small tooth each. Pharynx

muscular about 14 — 18 % Of total body length. Procorpus muscular, gradually

expanding to form an ovoid, median bulb. Isthmus broad expanding slightly to form

basal bulb. Anterior pharynx muscular, 55 — 61 % of pharyngeal length. Nerve ring encircling isthmus almost at its middle. Hemizonid conspicuous, slightly posterior to

the nerve ring. 73 — 85 % of the total pharyngeal length. Excretory pore posterior to hemizonid.

74 Reproductive system monoprodelphic, Ovary reflexed laterally or ventrally.

Oocytes arranged in several rows. Oviduct long, narrow gradually expanding to form

a spermatheca. Spermatheca set-off from the uterus by a constriction. Uterus well

developed, divisible into glandular and a short muscular part. Vagina muscular. Vulva

posterior with circular pore-like opening. Post-uterine sac short about 0.5 - 0.8 vulval

body diam. long. Rectum 1.1 to 2.0 anal body diam. long. Phasmids distinct. Tail

long, filiform, 2.7 to 5.0 times vulva-anus distance.

Males: Body smaller than females. Anterior end similar to that of females except for

four cephalic setae. Testis reflexed, flexure on right side of intestine. Spicules arcuate, cephalated, curvature more at the proximal end, distal tip curved anteriorly, 1.6 — 1.9 anal body diam. long. Gubernaculum broad, keel-like with rounded base, proximally notched, distal end with a fine tip, 42 — 50 % of spicules length. Genital papillae 9 pairs; 3 pairs precloacal, 6 pairs postcloacal. Genital papillae formula: vi, v2, v3d / v4, ad, phasmids, (v5. v6. v7), pd. vi almost at the level of spicule head, v4 at the level of lower cloacal lip. Tail divisible in two parts; a short conoid part and a long filamentous part.

Type habitat and locality: Farmyard manure collected from Aligarh, U.P., India.

Type specimens

Holotype: Female on slide Diplogastrellus curvatus sp. n./8 deposited in the nematode collection of the Department of Zoology. Aligarh Muslim University, Aligarh.

Paratjpes: Eight females and ten males on slides Diplogastrellus curvatus sp. n./1-7 deposited in the nematode collection of Section of Nematology, Department of

Zoology. Aligarh Muslim University, Aligarh.

Diagnosis and relationships

Diplogastrellus cur vatus sp. n. is characterized by small body, sharp and anteriorly directed dorsal tooth supported by a small tooth, a short post-uterine sac.

75 amphidial apertures at the base of cheilostom, arcuate and cephalated spicules with

anteriorly curved distal tip, and a broad, keel-like gubernaculum proximally with a

notch and a fine tip distally and nine pairs of genital papillae.

The new species resembles Diplogastrellus cerea Kiontke & Sudhaus, 1996 in

body length, tail length, in b value in female, in general shape of gubernaculum and

number of genital papillae. However, the new species can be differentiated from D.

cerea in having more slender body (a 35.1= — 42.9 vs 18.5 — 27.3), in smaller body

width (14 — 18 µm vs 20 — 43µm), slightly smaller pharynx (89 — 103µm vs 107 —

131µm in females and 83 — 97 pm vs 103 — 120 pm in males), anteriorly located

vulva (V=42-55% vs 61 —69 %), slightly longer tail (c = 2.1 — 3.0 vs 3.4-5.2), in having greater c' value (21.9 — 33.9 vs 8.9 — 15.0), structure of post-uterine sac (not bilobed vs bilobed), shape and length of spicules (more strongly ventrally curved in the proximal region with curved distal tip, 18 — 21 pm long vs uniformly curved ventrally, distal tip not curved, 24 — 28 pm long), in shape and length of gubernaculum (broader, distally with a fine tip, caudal margin continuous or entire and 8 — 10 pm in length vs relatively narrower. roundish, distally with a groove, caudal margin interrupted and 12 — 14 µm in length) and in position of genital papillae (v I at the level of spicule head when retracted. ad far anterior to v5 and pd posterior to v7 vs vi beyond the spicular range when retracted, ad close to GP v5 and pd and v7 at the same level).

Etymology: The species is named after the curved tip of the spicules.

76 T-qoo~

Table 5: Measurements (in µm) of Diplogastrellus curvatus sp. n. Mean and S.D. given in parenthesis.

Characters Holotype Females (n = 8) Males (n = 10) L 550 530-643 (590.4 f 39.3) 455-514 (487.3 f 21.0)

a 36.67 35.1 - 42.9 (38.5±2.6) 36.1 - 42.6 (38.5±2.1)

b 6.04 5.5-7.1(6.3±0.6) 4.9-6.1(5.5±0.3) c 2.24 2.1-3.0(2.4±0.3) 2.3-2.7(2.5±0.1)

c' 27.22 21.9-33.9(27.6±4.0) 15.6- 19.1 (16.8±0.9)

V 45.09 41.7-54.9(46.7±3.9) T 19.8-28.4 (24.6 ± 2.6) Maximum body width 15 14- 18 (15.4 ± 1.1) 12 - 14 (12.7 ± 0.8) Lip width 5 5-6(5.5±0.5) 4-5(4.8±0.4) Length of stoma 10 9- 11 (10.3 ± 0.7) 9- 10 (9.5 t 0.5) Pharynx 91 89 -103 (94.4 ± 4.3) 83 - 97 (83.4 ± 4.4) Excretory pore from anterior end 79 73 - 85 (79.0 ± 3.7) 74 - 82 (77.6 ± 2.5) Hemizonid from anterior end 74 70-82 (75.1 ± 3.7) 70-79(73.1 ± 3.1) Nerve ring from anterior end 68 63 - 71 (68.4 ±2.4) 60 - 68 (64.7 ± 2.4) Median bulb (length) 13 10- 12 (11.0 ±0.7) 9- 10 (9.8 ± 0.4) Median bulb (width) 8 8 - 10 (8.8 ± 0.8) 6 - 7 (6.9 ± 0.3) Pharynx base to gonad 97 70-115(93.0±14.4) 50-100(81.0±14.6) Anterior gonad 60 60 - 119 (86.9 ± 20.7)

Post uterine branch 9 8- 15 (10.6 t 2.3) VBD 15 14-18(15.6±1.1)

Vulva 248 248 - 291 (274.3 ± 12.0) Vulva - anus distance 57 57-70 (63.6 ± 3.9) Rectum I1 11-16(13.0±1.6) 1 12-15(12.8±0.98) Tail 245 175-305 (252.5 ± 41.98) 180-210 (196.7 ± 8.1) ABD 9 8-10(9.1±0.6) I1-12(11.7+0.5) Phasmids from anus/cloaca 14 14-19(17.5±2.1) 12-19(16.0±1.8) Testis 90 - 145 (120.2 ± 14.9) Spicules along axis 18 - 21 (19.8 ± 0.98) Spicules along cord 15 - 17 (16.2 ± 0.6) Gubernaculum 8 - 10 (9.1 ± 0.5)

77 A /r

j F

H

11

D

I— - 20ti

Fig. 9. Dip/ogustrellus curL'utus sp. n. A. Entire tmaie, B. Entire male, C. Pharyngeal region. D. Anterior region, F. Female Reproductive tract (Anterior). F. Female posterior region. G. Male posterior region. II. Spicules & Gubernaculum.

78 Fig. 10. Diplugasirellw curt&inus sp. n. A. 13. Anterior region showing stoma. C. Pharyngeal region. D. Female reproductive tract. E. Vulval region, F. Female posterior region, G. II. Male posterior region showing spicules & gubernaculum. (Scale bars - 2011n). Diplogastrellus ueograeiloides sp. n. (Fig. 1l, 12)

Measurements: In Table 6.

Descriptions

Females: Body almost straight upon fixation. Cuticle with fine transverse and

longitudinal striations. Amphidial openings oval, far below the base of stoma. Lip

region round and continuous with body contour. Lips fused, each with a papilla.

Stoma about 1.7 to 2.2 lip-widths long. Cheilostom euticularized, slightly longer than

wide, cheilostomal flaps extending up to labial contour. Gymnostom anisotopic,

dorsal wall smaller than suhventrals. Stegostom anisotopic and anisomorphic. Dorsal

tooth large with ventrally directed apex. Posterio-ventral aspect strongly cuticularised,

smooth. Anterio-dorsal aspect not cuticularised. Subventrals with a small pointed

tooth each directed dorsally. Procorpus muscular, median bulb elongate, about 2 to

2.6 as long as wide. Isthmus long, almost of uniform diam., gradually expanding to an ovoid basal bulb. Anterior pharynx 62 — 65 % of total pharyngeal length, Nerve ring

in anterior half of isthmus, generally just posterior median bulb. Hemizonid almost at

level of nerve ring. Excretory pore posterior to hcmizonid, 69 — 79 % of pharynx length. Cardia conoid. Intestinal lumen wide.

Reproductive system monoprodelphic with reduced posterior branch. Anterior branch on right side and posterior reduced branch on left side of intestine. Ovary reversed. Oocytes arranged in one or more rows, Oviduct narrow, gradually expanding into a spermatheca. Spermatheca slightly set off from uterus by constriction, containing sperms. Uterus divisible into long glandular and a short muscular parts. Vagina short, muscular. Vulva) opening circular, 323 — 525 Pm from the anterior end. Post-uterine sac present about 1.6 — 2.3 vulva[ body diam. long.

Lumen of post-uterine sac wide, containing sperms and degenerated oocytes-like

80 structures representing the remnants of the ovary. Rectum about one to one and a half

anal body diam. long. Phasmids 2 — 3 anal body diam. posterior to anus. Tail long,

filiform, whip-like, 1.6-2.8 times vulva-anus distance.

Males: Body smaller than females, slightly curved in posterior region on relaxation.

Anterior end similar to that of females except for four cephaplic setae. Testis ventral.

reflexed, flexure on right side. Spicules stout, ventrally arcuate with blunt tips, 1.3 —

1.6 anal body diam. long. Gubemaculum somewhat globose, with a small distal

sleeve. 42 -- 52 % of spicules length. Genital papillae eight pairs; three pairs

precloacal and five pairs postcloacal. Genital papillae formula: vi, v3d, v2/ v4, ad,

phasmids, (v5, v6), pd. vi beyond the spicular range and v4 far posterior to cloaca

slightly anterior to ad, v7 absent. Tail long filiform, 18 — 24 times anal body diam.

long.

Type habitat and locality: Farmyard manure collected from the District Baramulla,

Kashmir, I&K.

Type specimens

Holotype: Female on slide DiplogastreUvs neograciloides sp. n./6 deposited in the nematode Collection, Department of Zoology, Aligarh Muslim University, Aligarh,

India.

Paratypes: Ten females and ten mates on slides Dplogastrellus neograciloides sp. n./1-5 deposited in the nematode collection of Department of Zoology, Aligarh

Muslim University, Aligarh, India.

Diagnosis and relationship

Diplogestrellus neogracdoides sp. n. is characterized by a slender body, far posteriorly located amphidial openings, a large dorsal tooth with ventrally directed apex, an elongate median bulb, post-uterine sac with remnants of gonad, stout and

81 arcuate spicules with blunt tips, globular gubernaculum with a distal sleeve and eight

pairs of small genital papillae, v4 far posterior to cloaca, v7 absent.

The new species resembles D. graciloides (Skwarra, 1921) Paramonov, 1952

in general morphometrics. However, the new species differs from D. graciloides in

shape and length of spicules (distal end blunt, 1.3 — 1.6 anal body diam. vs pointed

distal end and I anal body diam. long), shape of gubernaculum (no indentation on

dorso-anterior side, distal sleeve present vs indentation on dorso-posterior side, sleeve

absent) and in number and arrangement of genital papillae (eight pairs: 3 pairs

precloacal, 5 pairs postcloacal vs nine pairs: 4 pairs precloacal, 5 pairs postcloacal).

Etymology: The name of the species is derived from its closely related species D. graciloides.

82 Table 6: Measurements (in µm) of Diplogastrellus neograciloides sp. n. Mean and S.D. given in parenthesis.

Characters Holotype Females (n = 10) Males (n = 10) L 886 773 - 1138 (945.2 ± 99.8) 765-948 (860.2 t 51.8) a 42.19 38.7-51.6(44.3±3.5) 41.2-52.7(46.6±3.2) b 4.76 4.5-5.7(5.17±0.4) 4.5-5.2(5.0±0.2) c 3.16 2.24-3.2 (2.7 ± 0.3) 2.5-2.9 (2.7 ± 0.1)

c' 23.33 20.8-41.8(27.7±5.7) 18.1-24.3(21.8±1.9) V 48.19 39.3-48.2(44.1 ± 2.8) T 28.0-35.5 (33.2 ± 2.3) Maximum body width 21 20-24(21.3±1.3) 17-21(18.5± 1.3) Lip Width 5 5-6(5.6±0.5) 4 - 6 (4.8 ±0.6) Length of stoma 11 10-11(10.6±0.5) 9-11(10±0.5) Pharynx 186 158-200(182.5±10.96) 156-193(172.1±9.7) Excretory pore from anterior end 137 123-147(135.2±6.97) 115-145(129.10±8.12) Hemizonid from anterior end 128 108 - 132 (122.4 ± 7.2) 105 - 126 (114.7 ± 5.9) Nerve ring from anterior end 125 105-129(119.9±7.5) 102 - 123 (111.5 ±5.6)

Median bulb (length) 30 24 -31 (28.7 t 2.3) 25 - 30 (26.9 ± 1.4)

Median bulb (width) 13 10 - 15 (12.7 t 1.2) 10-13(10.8±0.6) Pharynx base to gonad 111 95 -20(132.3 ± 30.2) 63-91(78.0 ± 9.1)

Anterior gonad 1 130 70-130(101.4± 18.9)

Post uterine branch 38 31 - 38 (35.9 t 2.1) VBD 20 16-24(20± 1.9)

Vulva - anus distance 179 152 - 223 (172 f 19.8) Rectum 18 15-18(16.5±0.8) 17-19(17.6±0.7)

Tail 280 270-460 (357 t 58.3) 280-360 (323.5 ± 25.7) ABD 12 11-16(13±1.3) 14-16(14.9±0.7) Testis 225 - 315 (286.4 ± 30.2) Spicules along axis 20 - 24(20.9 ± 1.2) Spicules along cord 18-21(19.3 ± 0.9) Gubemaculum 9 - 11 (9.6 ± 0.7)

83

~7

G

E 0 N

F

A, B'--y 5O ttm : .

C 20 inn

Fig. 11. Diplogastrellus neugrot•iloMt's sp. n. A. Entire female, B. Entire male, C. Anterior region. U. Pharyngeal region, F. Female Reproducti e tract (Anterior). F. Female posterior region, G. Male posterior region.

8.1

B D C

44 ,

> t Ii))

r •',

F , 1 1 .. I

•

J K 7 rI aJ Fig. 12. Diplogastrellus neograciloide.s sp. n. A. B. Anterior region sho\% inu stoma. C. Anterior pharynx, 1). Median bulb, F. F. Post-uterine sac, G. Reflexed testis, H. Female posterior region, I-K. Male posterior region showing spicnles & gubernaculum, (Scale bars 20pun). Diplogastrellus kashmeriensis sp.n. (Fig. 13, 14)

Measurements: In Table 7.

Descriptions

Females: Body slender, small tapering towards both extremities. Cuticle with fine

transverse and longitudinal striations. Lip region continuous with body contour. Lips

fused, each provided with a papilla. Amphidial apertures circular, at the level of

dorsal tooth. Stoma longer than wide. Chilostomal walls cuticularised, cheilorhabdial

flaps extending beyond labial contour. Gymnostomal walls anisotopic, dorsal wall

shorter than the subventrals. Stegostomal walls anisomorphic. Dorsal wall with a

large tooth directed anteriorly supported by a smaller tooth at base. Subvenlrals with

small teeth. Procorpus muscular, median bulb ovoid. Isthmus long, gradually

expanding posteriorly to form an ovoid basal bulb. Anterior pharynx 49 — 59 % of

total pharyngeal length. Nerve ring in the anterior half of isthmus about 61 — 82 % of

pharyngeal length. Hemizonid opposite to the nerve ring. Excretory pore slightly

posterior to hemizonid about 66 — 94 % of total pharynx length. Cardia distinct.

Intestinal lumen wide.

Reproductive system monoprodelphic, on the right side of intestine. Ovary

long reversed, oocytes arranged in several rows. Oviduct long, slender, gradually expanding to form a spermatheca. Spermatheca separated from uterus by a constriction, usually containing spermatozoa. Uterus divisible into a short muscular and a long glandular part. Vagina muscular. Vulva) opening oval. Post-uterine sac present, 0.8 — 1.3 vulva) body diam. long, on the left side of intestine. Lumen of post- vulval sac wide, containing glandular structures in the distal or posterior part.

Phasmdial apertures large, 1.1 — 1.6 anal body diam. posterior to anus. Tail long, filiform, 2.5 to 3.4 times vulva-anus distance,

86 Males: Body smaller than females in fixed specimens, curved posteriorly. Anterior

region similar to that of females except for four cephalic setae. Testis ventral,

reflexed. Spicules slender, arcuate, 1.6 to 1.9 times anal body diam. long.

Gubernaculum slender, with a large sleeve, 44 — 52 % of spicules length. Genital

papillae nine pairs: three precloacal and six postcloacal. Genital papillae formula: (vI,

v2), v3d / v4, ad, phasmids, (v5, v6, v7), pd. GP vi and v2 close to each other and within the spicular range. GP v4 at the lower cloaca) lip. Tail about 8 — 14 times anal body diam. long and divisible in two parts: a short conoid part and a long filamentous part.

Type habitat and localit} • Farmyard manure collected from Baramulla, Kashmir,

J&K.

Type specimens

Holotype: Female on slide Diplogastrellus kashmeriensis sp. n./I I deposited in the nematode Collection. Department of Zoology, Aligarh Muslim University, Aligarh,

Paratypes: Ten females and ten males on slides Diplogastrellu kashmeriensis sp.n./l-

10 deposited in the nematode collection of Department of Zoology, Aligarh Muslim

University, Aligarh.

Diagnosis and relationship

Diplogastrellus kashmeriensis sp.n. is characterized by a small body, tubular stoma, claw-like dorsal tooth and a supporting tooth, presence of post-uterine sac; slender gubernaculum with a large sleeve and nine pairs of genital papillae.

The new species resembles D. monhvsteroides (Butschli, 1874) Paramonov,

1952 in body size, morphometric values and female reproductive system. However, the new species can be differentiated from D. monhysteroides in the shape of gubemaculum (slender with a large sleeve vs shoe-like without a distal sleeve) and the

87 number and arrangement of caudal papillae (9 pairs; 3 precloacal, 6 postcloacal vs 12 pairs; 3 precloacal, 9 postcloacal).

Etymology: The name of the species is derived from the name of the place from where it was collected.

88 Table 7: Measurements (in gm) of Diplogastrellus kashmeriensis sp. n. Mean and S.U. given in parenthesis.

Characters Holotype Females (n = 10) Males (n = 10) L 577 498 - 688 (575.9 t 62.2) 404 - 543 (470.2 ± 38.2) a 28.85 25.5 - 34.4 (28.7 f 2.4) 25.25 - 33.94 (29.40 ± 2.42) b 5.50 4.4 - 6.4 (5.3 ± 0.6) 4.08-6.20 (4.74 ± 0.62) c 2.96 2.8-3.4(3.0±0.2) 2.85 - 4.28 (3.35 ± 0.42) C' 19.50 13.2-20.8 (16.6 ± 2.7) 7.86-13.15 (10.42±1.66) V 53.90 52.5 - 58.8 (55.4± 2.0) T 32.18- 45.45 (40.31 t 4.36) Maximum body width 20 17-22(20.1 ± 1.4) 15- 17 (16.0 ± 0.45) Lip width 7 5- 8(7.0±0.9) 5 - 7 (5.90 ± 0.54) Length of stoma 10 9-12(10.4±0.9) 8-9(8.80±0.40)

Pharynx 105 79-124(109.7± 12.0) 79-115(100.10± 9.50) Excretory pore from anterior end 80 74 -91 (82.7 t 5.6) 69 - 80 (74.70 t 3.49) Hemizonid from anterior end 75 70 - 87 (78.0 t 5.6) 65 - 76 (70.30 t 3.5) Nerve ring from anterior end 66 65 - 83 (72.0 t 5.9) 60 - 72 (64.9 t 3.7) Pharynx base to gonad 89 70-100(85.7±11.0) 20-50(34.9± 11.2)

Anterior gonad 117 95-157(122.8± 16.6) Post uterine branch 21 15 - 25 (20.3 t 2.5) VBD 18 16-21(19.4±1.4) Vulva 311 279 - 365 (318.2 ± 27.4) Vulva - anus distance 68 60-73(66.1 ± 5.5) Rectum 20 12-20(16.9±2.3) 15-20(17.2±1.8)

Tail 195 150-250(191.3±31.7) 110-171(142.6±21.95)

ABD 10 10-13(11.611.0) 13-15(13.7±0.6) Phasmids from anus/cloaca 16 13- 17(14.2 f 1.3) 14 - 18 (15.8 ± 1.5) Testis 130-219(189.9±27.4) Spicules along axis 21 - 26 (23.5 ± 1.6) Spicules along cord ; 19 - 21 (20.4 ± 0.7) Gubernaculum 10-12(11.3±0.6)

89 ❑C

Fig. 13. Diplogastrellus kashmeriensis sp. n. A. Entire female, B. Entire male, C. Pharyngeal region, D. Female Reproductive tract (,Anterior), F. Post-uterine sac. F. Female posterior region, G. Male posterior region.

90 r• T' F C D E

f • .

G

K

Fig. 14. Diplogustrellus kashmerien si.s Sp. n. A. Anterior pharynx, B. Median bulb & posterior pharynx, C. Anterior region showing stoma (lateral). D. Anterior region showing amphids (dorsoventral). E. F. Female reproductive tract (Anterior branch), G. H. Post-uterine sac, I. Female posterior region, J. K. Male posterior region showing spicules & gubernaculum, (Scale bars 20µm). Diplogastrellus ioktakai sp. n. (Fig. 15, 16)

Measurements: In Table 8.

Descriptions

Females: Body small, tapering towards both the extremes, almost straight upon

fixation. Cuticle with fine transverse striations. Amphidial apertures oval, at the level

of dorsal tooth. Lip region continuous with body contour. Lips fused, cacti with a

labial papilla. Stoma barrel-shaped, 1.6 — 2.2 times as long as lip diam.

Cheilorhabdions cuticularized, plate-like. Oymnostom anisotopic, dorsal wall shorter

than the subventrals. Stegostom anisotopic and anisomorphic. Dorsal wall with a large

tooth with ventrally directed apex. The subventrals with a small tooth each. Pharynx

muscular about 13 — 17 % of total body length. Procorpus muscular, gradually expanding to form an ovoid, median bulb. Isthmus short expanding gradually to form a basal bulb. Anterior pharynx muscular, 55-67% of the pharyngeal length. Nerve ring encircling isthmus anteriorly at about 67 — 82 % of pharyngeal length. Hemizonid conspicuous, posterior to the nerve ring. Excretory pore posterior to hemizonid, almost opposite to basal bulb.

Reproductive system monoprodelphic, Ovary reflexed. Oocvtes arranged in two or several rows. Oviduct narrow gradually expanding to form a sperntatheca.

Spermatheca set-off from the uterus by a constriction, filled with sperms. Uterus divisible into a large and prominent glandular part and a small and narrow muscular part. Vagina muscular. Vulva opening circular, pore-like. Post-uterine sac short about

0.7 to 0.9 vulval body diam. long. Rectum 1.0 to 1.5 anal body diam. long. Phasmids distinct, 1.6 to 2.0 anal body diem. posterior to anus, Tail long, tiliform, 3.8 to 5.5 times vulva-anus distance.

92 Males: Body smaller than females. Anterior end similar to that of females except for

four cephalic setae. Testis reflexed, flexure on right side of intestine. Spicutes arcuate,

proximally knobbed and distally tapering to a fine tip, 1.4 — 1.8 anal body diam. long.

Gubemaculum 29 — 41 % of spicules length, keel-like, proximally with a notch and

distally with a pointed extension and a sleeve. Genital papillae 9 pairs; 3 pairs

precloacal, 6 pairs postcloacal. Genital papillae formula: vi, v3d, v2/ v4, ad,

phasmids, (v5, v6, v7), pd. vl anterior to the spicule head. v2 and v4 close to the

cloacal region, ad just anterior to v5. Phasmids between v5 and v6. Tail long,

filamentous.

Type habitat and locality: Cow drop collected from Khornapaat, Loktak Lake,

Manipur.

Type specimens

Holotype: Female on slide Diplogactrellu.s to/cm/cal sp. n./8 deposited in the nematode

collection of the Department of Zoology, Aligarh Muslim University, Aligarh.

Paratypes: Ten females and ten males on slides Diplogastrellus loktakai sp. n./1-7

deposited in the nematode collection of Section of Nematology, Department of

Zoology, Aligarh Muslim University, Aligarh.

Diagnosis and relationships

Diplogastrellus loktakai sp. n. is characterized by small body, large dorsal

tooth with ventrally directed apex, a short post-uterine sac, amphidial apertures oval, at the level of dorsal tooth, arcuate and proximally knobbed spicules, a keel-like gubernaculum proximally notched, distally with an pointed extension and nine pairs of genital papillae.

The new species resembles Diplogasirelhrs cerea Kiontke & Sudhaus, 1996 in having similar body length, same b value, same a value in males, pharyngeal length, in the shape and length of spicules, general shape of gubernaculum and number of

93 genital papillae. However, the new species can be differentiated from D. cerea in

having more slender body (a = 30.7-39.8 vs 18.5 — 27.3), stegostom with larger

dorsal tooth vs medium sized tooth, smaller c value (c — 1.9 — 2.4 vs 3.4 — 5.2 in

females and 2.3 — 3.0 vs 4.2 —6.1 in males), in having greater c' value (26.4-33.5 vs

8.9 — 15.0 in females and 13.9 —21.9 vs 4.6 — 9.1 in males), in smaller body width in males (16-19 pm vs 21 — 30 pm), anteriorly located vulva (V=42.2-51.1 % vs 61-

69 %), structure of post-uterine sac (not bibbed vs bibbed), longer tail in both the

sexes (290-335 pm vs 113— 241 pm in females and 180-285 gm vs 74 — 160 pm in males), bursal remnants (invisible vs clearly visible), length and structure of gubernaculum (7 —9 µm long with entire caudal margin, distal sleeve present vs 12-

14 pm with interrupted caudal margin, sleeve absent) and position of genital papillae

(v3d lies after v1, ad very close to or at the level of v5 and pd posterior to v7 ur v2 lies after vl, ad relatively more anterior to v5 and v7 and pd at the same level).

Etymolog': The name of the species is derived from Loktak Lake from which it was collected.

94 Table 8: Measurements (in pm) of Diplogaslrellus (oklakai sp. n. Mean and S.D. given in parenthesis.

Characters Holotype Females (n = 10) Males (n = 10) L 691 613-741(689.4±340) 513-641(566.9±42.8)

a 32.90 30.7-39.8(35.213.0) 28.5-35.5 (332 t2.0)

b 7.19 6.0-7.8(6.9±0.6) 5.3-6.8(5.810.5) c 2.09 1.89- 2.43 (2.2 t 0.2) 2.3-3.0(2.6±0.2)

c' 30.0 26.4-335(29.9±2.2) 13.9-21.9(16.9±2.4)

V 42.84 422 -51.1 (46.1 t 2.4) T 34.4- 40.5(37.4± 1.9) Maximum body width 21 16-22(19.5±1.9) 16- 19(17.1 ± 1.I) Lipwidth 6 5-6(5.7±0.5) 5-6(5.3±0.5) Length of stoma 10 8-11(10.2+0.9) 9-10(9.6 t 0.5)

Pharynx 96 85-111(101.0±6.9) 91-105(98.1±4.8)

Excretory pore from anterior end 90 74-95 (89.3 = 6.3) 80-91 (84.9 t 4.3) Hemizonid from anterior end 87 69-91(85.9± 6.7) 76-87 (80.9 t 4.1) Nerve ring from anterior end 78 64-86(77.7 t 6.8) 69 -80(73.2 t 3.9) Median bulb (length) 14 12 - 15 (14.0 f 1.1) 11 - 14(12.4± 1.0) Median bulb (width) II 10-12(11.110.7) 9-10(9.8 f 0.4) Pharynx base to gonad 70 70-100 (84.0 f 8.5) 18-60(38.6 t 14.1) Anterior gonad 130 122-160(132.6± 11.0) Post uterine branch II 11-15(13.0 f 1.6) VBD 16 15-19(16.9±1.1)

Vulva 296 296-361 (317.6+ 17.5)

Vulva-anus distance 65 55-77 (67.3 +5.9)

Rectum 14 11-16(13.2+ 1.4) 13-16(14.9} 1.0)

Tail 330 290-335(313.5+ 15.3) 180-285(218.0±31.7)

ABD II 10-11(10.6+0.5) 12-14(12.9+0.8)

Phasmids fromanuslcloaca 20 17-20(18.911.0) 14- IS (16.2+ 1.1) Testis 180-235(212.2 f 21.7) Spicules along axis 19-24 (21.3 1.4) Spicules along cord 16-20(18.1 s 1.1)

Gubemaculum 7-9(7.6 ± 0.7)

95 A

F

I

E

N O

LIIi

Fig. 15. Diplogastrellu.s loklakai sp. n. A. Entire female, B. Entire male, C. Anterior region showing amphids dorsoventrall . 1). Pharyngeal region. F. Female Reproductive tract (Anterior), F. Female posterior region, G. Male posterior region, 11. Spicules & Guhernaculum.

96 I .L \1

E F

j 1 1

1L I 0 D H j\ •.':..

i I 1 4 I I

1•~P~ 1 U `~ G J

fig. 1(. I)i/?1uga.stre!lus loktakai sp. n. A. I'har\nueal region, B, C. Anterior region showing stoma, B. Female reproductive tract (Anterior branch), F. uterus with egg, F. Post-uterine sac, G. Female posterior region . H-J. Male posterior region showing spicules & gubernaculum. (Scale bars 20µm). Diplogastrellusgracilis (BUtschli, 1876) Paramonov, 1952 (Fig. 17, 18)

Measurements: In Table 9.

Description

Females: Body slender, almost straight upon fixation tapering at both extremities.

Cuticle with fine transverse and longitudinal striations. Lip region continuous with

body contour. Amphidial opening ovoid, post-labial. Stoma longer than wide.

Cheilostom arched with cuticularized walls. Gymnostomal walls anisotopic with dorsal vial! shorter than subventrals. Stegostom anisotopic and anisomorphic. Dorsal tooth massive, ventrally directed with serrated anterior margin. Each subventral wall

provided with one ridge. Pharynx about 14 - 20 % of total body length. Procorpus weakly muscular, median bulb oblong. Isthmus narrow, gradually expanding posteriorly to forth a glandular basal bulb. Nerve ring at about 71 - 83 % of pharynx length. Hemizonid posterior to the nerve ring, 82 - 93 % of the total pharyngeal length. Excretory pore located posterior to hemizonid, about 86-98% of pharyngeal length. Cardia present. Intestinal lumen wide.

Reproductive system monoprodelphic. Ovary reflexed on right side of the intestine. Germinal zone long with several rows of oocytes. Oviduct narrow, long gradually expanding to form spermstheca. Spermatheca not set-off from uterus, usually filled with sperms. Uterus divisible into a long, glandular often filled with sperms and a short muscular part. Vagina anteriorly directed. Body markedly narrow posterior to vulva. Post uterine sac absent. Vulva- anus distance 2.1 to 2.8 anal body diam. long. Rectum about 1.0 to 1.3 anal body diam. long. Phasmids 1.2 to 1.5 anal body diam. posterior to anus. Tail 14 to 17 anal body diams long, filifomt, gradually tapering to a fine terminus.

98 Males: Body smaller than female, strongly curved in posterior region. Testis single,

on right side of intestine and reflexed laterally. Spicules long, setose, about 2.7 to 3.5

anal body diam. long, strongly curved ventrally with distinct rounded capitula.

Gubernaculum with keel-like proximal end and lateral sleeves at distal end

surrounding terminal region of spicules. Eight pairs of genital papillae: 2 precloacal,

2 adcloacal and 4 postcloacal pairs. Genital papillae formula: vI, v3d, v2/ v4, ad,

phasmids, (v5, v6), pd. Tail filiform divided into short conical part followed by whip-

like posterior part.

Habitat and locality: Chicken manure collected from Haldwani, Uttrakhand.

Voucher specimens

Ten males and ten females on slides Diplogastrellus gracilis/t -5 deposited in

the nematode collection of Department of Zoology, Aligarh Muslim University,

Aligarh.

Re narks

Measurements and description of our specimens conform well with that of the original specimens described by (BUtschli, 1876) Paramonov, 1952. However, differences were observed in total length of the body in both the sexes (0.7 — 0.99 mm vs 1.35 — 1.39 min in females and 0.6 — 0.8 mm vs I — 1.07 mm in males), smaller V value (62.6 — 71.3 vs 73 — 77), smaller a value in males (25.3 — 29.1 vs

36.0), smaller b value in males (4.9 — 6.7 vs 8.2), smaller c value in males (4.2 — 6.5 vs 7.1), presence of massive dorsal tooth with serrated anterior-ventral margin vs smaller tooth lacking serrations), bursal rudiments (absent vs present), position of v 1

(within the spicular range when retracted vs beyond the spicular range). Table 9: Measurements (in µm) of Diplogastrellusgracf/rs. Mean and S.D. given in parenthesis.

Characters Females (n = 10) Males (n = 10) L 655-997 (830.1 f 89.2) 559-865(731=109)

a 23.5-34.1 (29.0 f 2.98) 25.3-29.1(27.4+1.1)

b 5.1-7.2 (6.3 t 0.6) 4.9- 6.7(5.8+ 0.5)

c 3.1-4.1 (3.6 t 0.3) 4.2-6.5 (5.3t 0.7)

c' 14-16.9(15310.9) 5.4-8.4(6.910.8)

V 62.6-71.3 (67.7+2.4)

T 43.5-57.7(49.9'- 5.0) Maximum body width 25-34(28.5±3.4) 20-32 (26.8 t 4.4) Lip width 7-9(7.8±0.8) 6-8(6.8;0.6)

Length of stoma 9-12(11 t 1.0) 9-11(102±0.6)

Pharynx 120-140(131.0 t 5.5) 107-139(125.9 t 10.4) Excretory pore from anterior end 113- 125(121.1 x 3.4) 99- 126 (114.7 t 8.99)

Hemizonid from anterior end 106-121 (114.8 t 4.4) 92-121(106.9 t 9.3) Nerve ring from anterior end 95-110(10].4 f 4.7) 83-105(95.3 t7.4) Median bulb (length) 20-30(23.5 f 2.6) 16-23(20.4+2.0)

Median bulb(width) 13-19(15.1±1.8) 10-15(13.3+2.0) Pharynx base to gonad 135 - 198(168.8 f 20.7 60-138 (93.1 * 23.7) Anterior gonad 137-345(263.9± 582) VBD 23-31(26.7±3.1) Vulva 410-682 (563.7 As 75.1) Vulva - anus distance 30-45 (36.2 f 3.9) Rectum 15-20(17* 1.7) 15-23(18.8±2.2) Tail 210- 270(230.2 t 16.5) 125- 165 (139.0 t 12.8) ABD 13-16(14.9 r0.8) 16-23(20.232.1)

Phasmids from anus/cloaca 18-20(19.5±0.7) 15-18(16.8+1.2) Testis 245 - 470 (376.0 t 76.1) Spicules airing axis 55 - 65 (60.7 i 3.8) Spiculcs along cord 45 - 55 (50.4 t 2.9) Gubereaculum 20-26(23.6± 19) L [

100

B C

G

. • : IF \. p. •\ Gi 20 gin J .1

______!J I), F p— —s 50 pill 6'

\ /1'

44

Fig. 17. Diploga.stre!lus grad/is. A. Entire female, B. Entire male, C. Pharyngeal region. 1). Female Reproductive tract (Anterior), E. Female posterior region, F. G. Male posterior region.

101

0 B C

F

E

Fig. 18. Diplogastrellus grad/is. A. Anterior phar\nx. B. C. Anterior region showing stoma. D. Vulval opening (dorsoventral). F. Female posterior region showing vulval region. rectum & anal opening. V. Male posterior region showing spicules & gubernaculum. (Scale bars - 20µm). Pseudacrostichus gen. a.

Diagnosis: Diplogastridae. Body small in size, gradually tapering towards both

extremities. Cuticle with prominent longitudinal striations. Lip region smooth, round

and continuous with body contour. Labial papillae and cephalic papillae in males

minute. Stoma longer than wide. Cheilostom well-developed. Gymnoslom anisotopic.

Stegostom anisomorphic with a large sharp dorsal tooth supported by a small tooth at

its base and a small tooth on each subventral wall. Corpus muscular. Median bulb

ovoid. Isthmus short expanding gradually to form a basal bulb, Cardia present.

Female reproductive system arnphidelphic. Ovaries reversed. Spicules arcuate,

cephalated. Guhemaculum large and massive, 50 — 60 % of total spicule length.

Genital papillae nine pairs. Three pairs precloacal and six pairs posteloacal. First two

papilla closely placed. Female tail long filiform and male tail divisible into a conoid

and a filamentous part.

Etymology. The name of the genus is derived from Acrostichus.

Type and only species: Pseudacrostichus musai sp. n,

Relationships

The new genus Pseudacrostichus gen. n. can be distinguished from all other

genera of Diplogastridae by having thick cuticle with strong longitudinal striations;

anisotopic gvmnostom with dorsal wall shorter than the subventrals, anisomorphic

stegostom with large and sharp anteriorly pointed dorsal tooth supported by a small

tooth at base, subventrals with a small tooth each; amphidelphic gonad with posterior

branch shorter; set off and slightly inflated spermatheca; large gubernaculum with a

thread like proximal extension. However, it is related to the genus Acrostichus, Rahm

1928 in having smooth and round lip region, anisotopic and anisomorphic stegostom, a large anteriorly pointed dorsal tooth, muscular pharynx, oval median bulb, amphidelphic female reproductive system, number of genital papillae, in position of

103 genital papillae vi and v2 (closely placed), arcuate spicules and massive

gubemaculum. However, the now genus can be differentiated from Acrostichus by

having anisotopic gymnostom, presence of stronger longitudinal striations, presence

of a supporting tooth at the base of dorsal tooth, lack of vaginal pouches, set-off

spermatheca and presence of a thread-like entension at the proximal end of

gubemaculum.

The new genus also resembles the genus Drplogasirellus, Paramonov, 1952 in

having a small supporting tooth at the base of dorsal tooth, anisotopic gymnostom, in

having more straight and pointed dorsal tooth. pharynx, ovoid median bulb and in

number of genital papillae. However, the new genus differs from Diplogastrellus in

having thicker cuticle with strong longitudinal striations, amphidelphic female

reproductive system, set-off spermatheca, presence of larger, well built or massive

gubemaculum and in the arrangement of genital papillae.

Pseudacrostichus musai sp. n. (Fig. 19, 20)

Measurements: In Table l0.

Descriptions

Females: Body small about 0.5 — 0.7 nun in length, almost straight upon fixation.

Body tapering towards both extremities. Cuticle with strong longitudinal ridges and fine transverse striations. Amphidial apertures oval, at the base of cheilostom almost at the anterior tip of dorsal tooth. Lip region continuous with body contour. Lips fused, each with a small labial papillae. Stoma harrel-shaped, longer than wide, 0.9-

1.3 times as long as lip diam. Cheilorhabdions well developed, cuticularized.

Gymnoslom anisotopic, dorsal wall shorter than the subventrals. Stegostom anisotopic and anisomorphic. Dorsal wall with a sharp, anteriorly pointed tooth supported by a

104 small tooth at base. The subventrals with a small tooth each. Pharynx muscular about

19 — 24 % of the total body length. Procorpus muscular, gradually expanding to form

an ovoid, median bulb. Median bulb ovoid, 19 — 22 pm long and 11 — 15 µm wide.

Isthmus short expanding posteriorly to form a glandular basal bulb. Anterior pharynx

67-69 % of total pharyngeal length. Nerve ring encircling at middle of the isthmus,

50-84 % of total pharyngeal length. Hemizonid slightly posterior to the nerve ring,

at about 83 % of the total pharyngeal length. Excretory pore further posterior to

hemizonid, at about 94 % of total pharyngeal length. Cardia distinct. Intestine with

wide lumen.

Reproductive system didelphic, amphidelphic, anterior gonad on the right side

and posterior gonad on the left side of the intestine. Posterior branch is always shorter

than anterior branch. Ovary reversed. Oocyaes arranged in several rows. Oviduct

narrow gradually expanding to form a spermatheca. Spermatheca slightly inflated, set-

off from the uterus by a constriction. Uterus well developed divisible into long

glandular and a short muscular part. Vagina muscular with circular pore-like opening.

Vulva-anus distance 4.97-6.4 times vulval body diameters long. Rectum 1.4 to 2.0

anal body diams long. Phasmids distinct, 0.9 — 1.3 anal body diameter posterior from

anus. Tail filiform, gradually tapering to a finely pointed tip, 0.7 — 1.0 times vulva-

anus distance.

Males: Body slightly smaller than females. Anterior end similar to that of females except for four cephalic setae. Testis reflexed, flexure on right side of intestine.

Spicules slightly arcuate, proximal half is broader than the distal half, cephalated, 1.4

— 1.9 anal body diam. long. Gubemaculum large, well built, 50 — 60% of spiculcs length, proximally with a thread-like extension, gradually tapering towards distal end.

Genital papillae 9 pairs; 3 pairs precloacal, 6 pairs postcloacal. Genital papillae formula: (vi, v2), v3d / v4, ad, phasmids, (v5, v6, v7), pd. GP vl and v2 placed close

105 to each other and within the spicular range. GP v3d at the level of v2. GP v4 slightly

posterior to lower cloacal lip. Phasmids posterior to ad GP v5, v6, v7 gouped

together at the junction of conoid and filiform parts of tail. GP pd opposite to v5-7

group. Tail in two parts; a short conoid part and a long filamentous part.

Type habitat and locality: Decayed banana rhizome collected from Haldwani,

Uttrakhand, India.

Type specimens

Ho!orype: Female on slide Pseudacrostiehus musai sp. n./6 deposited in the nematode collection of the Department of Zoology, Aligarh Muslim University, Aligarh.

Parac pes: Ten females and ten males on slides Pseudacroslichus musai sp. nil-5 deposited in the nematode collection of Section of Nematology, Department of

Zoology, Aligarh Muslim University, Aligarh.

Remarks

The new species Pseudacrosticiovs musai sp. n. is characterised by presence of strong longitudinal striations; oval amphidial openings present at the base of cheilostom or at the anterior tip of dorsal tooth; barrel-shaped stoma, anisotopic gymaostom with slightly shorter dorsal wall than the subventrals, anisonvorphie stegostom with large and sharp anteriorly pointed dorsal tooth supported by a small tooth at base, subventrals with a small tooth each; amphidetphic gonad with posterior branch shorter than the anterior one in all the specimens; set off spermatheca, inflated and separated from oviduct and uterus; slightly arcuate, cephalated spicules with proximal half broader than the distal half, a large, and well built gubernaculum with a thread-like extension proximally and gradually tapering towards the distal end, nine pairs of genital papillae; 3 pairs precloacal, 6 pairs postcloacal; GP vl and v2 grouped together above the cloaca, GP v5, v6, v7 grouped together at the junction of conoid and filiform parts of tail.

106 Etymology: The name of the species is derived from the name host tree from which it was collected.

107 Table 10: Measurements (in µm) of Pseudacrostichus musai sp. n. Mean and S.D. given in parenthesis.

Characters Holotype Females (n = 10) Males (n = 10) L 589 536-666 (599.9 ± 48.1) 476-657 (549.7 f 55.4) a 19.63 17.7-23.9(20.98± 1.7) 20.1-26.3(22.7± 1.9) b 4.43 4.1 - 5.4 (4.6 ± 0.4) 4.1 - 5.3 (4.5 ± 0.4) c 4.06 4.1-5.0(4.4±0.3) 4.1 -5.6(5.1 ± 0.4) c' 9.67 7.9 - 11.5 (9.6 ± 1.0) 4.8 - 7.4 (5.6 ± 0.7) V 48.39 48.4-53.4(50.4± 1.4) T 43.97 - 54.7 (48.2 ± 3.2) Maximum body width 30 23 - 37 (28.9 ± 4.3) 20 - 30 (24.3 ± 2.7) Lipwidth 10 8-10(9.5±0.7) 8-10(8.9±0.7) Length of stoma 10 9-11(10±0.6) 9-10(9.5±0.5) Pharynx 133 119-141(129.6±5.9) 115-125(121.3±3.6) Nerve ring from anterior end 105 97 - 111 (103.4 f 3.6) 89 - 103 (98.1 t 4.0) Median bulb (length) 21 19-22 (20.6 ± 0.9) 18-20 (19.4 t 0.7) Median bulb (width) 12 11 - 15 (12.6 ± 1.4) 10- 13 (11.4 t 0.8) Pharynx base to gonad 55 55-126(80.9±20.4) 40-71(53.4± 10.0) Anterior gonad 97 75-115(91.5±15.96) Posterior gonad 71 50 - 83 (64.3 ± 11.7) VBD 28 23 - 36 ( 28.4 ± 4.5) Vulva 285 275 - 336 ( 302 ± 22.1) Vulva-anus distance 159 130-187 (162.4 ± 19.1)

Rectum 25 22 - 29 (25 ± 2.1) 22 - 25 (24.1 f 1.0) Tail 145 110-150(135.5±12.5) 95-125(108.5±10) ABD 15 13-16(14.2±1.2) 17-23(19.6±1.7)

Phasmids from anus/cloaca 13 11 - 17 (14.1 t 1.96) 15-20(16.7±1.6) Testis 220 - 350 (266.5 ± 44) Spicules along axis 30-32 (30.7 ± 0.8) Spicules along cord 26 - 29 (27.4 ± 1.1) Gubemaculum 16- 18 (16.8 ± 0.8)

108 1111/

rV 9

,"®

l

Fig. 19. Psc'udacrosticl:us tnusui sp. n. A. Entire ftmale, B. Entire male, C, D. Anterior region. F. Pharyngeal region, F. Female Reproductive tract (Anterior). G. Female posterior region, 1-1. Male posterior region, I. Gubernaculum.

109 A B C F

i w ~ 3.

E V.

0

i* `t

fl

Fig. 20. Pseudacruiichw mtesai sp. n. A. Anterior region shoe ing stoma, 13. Anterior region showing cuticle. C. Anterior region showing amphids (dorsoventral), I). Pharyngeal region. L. Cuticle showing longitudinal ridges, F. Female reproductive tract (anterior branch), G. Vulval region, II. Female posterior region. I. J. Male posterior region showing spicules & gubernaculum. (Scale bars - 20µm). Subfamily Demaniellinae Paramonov, 1951

Diagnosis: Cheilostom and gymnostom somewhat uniformly long. Stegostom with a

small tooth, not protrusible. Median bulb twice as long as its width, narrow

posteriorly. Ovaries paired or unpaired. Bursa absent.

Type genus: Demaniella Steiner, 1914

Other genera: Metadiplogaster Mel-1, 1961

Genus Demaniella Steiner, 1914

Diagnosis: Body 0.6-1.2 mm long. Lips six with very small papillae. Ventral

gymnostomal wall noticeably longer than that of dorsal, tube-like. Dorsal swelling of

stegostom with a plate-like, immovable tooth. Pharynx slender, median bulb oblong,

twice as long as wide. Female reproductive system paired. Gubernaculum long,

slender. Tail in both sexes very' long, filiform.

Type species: Demaniella cibourgensi.s Steiner, 1914

Demanie11a keirakensis Mahamood, Shah & Ahmad, 2006

(Fig. 21, 22)

Afeasurement.s: In Table 11.

Descriptions

Females: Body almost straight on relaxation, tapering towards both extremities. Body

length ranged between 0.76 - 0.99 mm. Cuticle with transverse and longitudinal striations. Lip region continuous with body contour. Lips six, the apex of each lip

with a fine process. forming a spine-like structure in lo\\ magnification. Labial

papillae small. Lateral fields with two lines. Amphidial apertures oval, just below the stegostom. Stoma longer than wide. Cheilostom weakly cuticularized, cheilorhabdial

flaps indistinct. Gymnostom short, anisotopic, dorsal wall shorter than subventrals.

Stegostum anisomorphic. Dorsal wall with a ventrally directed tooth. Posterio-ventral

aspect of tooth more sclerotized than anterio-dorsal aspect. Each subventral with a

small ridge. Procorpus muscular, median bulb elongate, 30 —40 µm long and II — 16

pm wide or 2.5 to 3.0 times as long as wide, Isthmus gradually expanding to basal

bulb. Basal bulb not clearly demarcated from isthmus. Anterior pharynx 69 — 75 % of

total pharyngeal length. Nerve ring encircling isthmus anteriorly, slightly posterior to

the median bulb, at 75 — 84 % of total pharyngeal length. Hemizonid at level of nerve

ring, 76 - 89 % of total pharyngeal length. Excretory pore posterior to hemizonid, at mid isthmus region, 82-93 % of total pharyngeal length. Cardia distinct. Intestinal lumen wide.

Reproductive system amphidelphic. Anterior branch longer, on right side; posterior shorter, on left of intestine. Ovaries reversed, oocytes arranged in several rows. Oviduct long, narrow, opening adaxially in the glandular part of uterus.

Spermatheca large, pouch-like, not continuous with oviduct anteriorly, containing sperms, set off from uterus. Uterus divisible into a bibbed glandular part and a short muscular part. Vagina muscular. Vulval opening circular. pore-like. Rectum 1.1 - 1.6 anal body diem. long. Phasmids distinct, 1.4 to 2.2 anal body diams posterior to anus.

Tail long, filiform, 0.7 - 1.0 times vulva-anus distance.

Males: Body smaller than females, anterior end similar to that of females except for four cephalic setae and posterior end curved on relaxation. Testes ventral with reflexed proliferation zone. Flexure on right side. Spicules arcuate. 1.4 to 2 anal body diams long. Gubernaculum slender, 56-74% of spicules length with a pair of flaps near distal tip. Distal tip spout-like, posteriorly directed. Genital papillae seven pairs;

112 two pairs precloacal, five pairs postcloacal. Genital papillae formula: vi, v2 / ad,

phasmids, (v5, v6, v7). pd. Genital papillae v3d and v4 absent. Right and left groups

of (v5, v6, v7) at different levels. Tail divisible into a short, conoid and a long,

filamentous parts.

Habitat and locality: Farmyard manure collected from Keirak, Thoubal, Manipur.

Voucher specimens

Nine females and ten males on slides Demaniella keirakensis/1-4, deposited

in the nematode collection of Department of Zoology, Aligarh Muslim University,

Aligarh.

Remarks

Measurements and description of our specimens conform vell with that of the

original specimens of Demaniella keirakensis described by Mahamood et al. (2006a)

in general morphometrics, presence of bibbed glandular part of uterus, long and

narrow oviduct, not connected with spermatheca anteriorly, opening adaxially in the

glandular part of uterus, presence of a large pouch-like spermatheca, in shape of

spicules and gubernaculum and number and arrangement of genital papillae. However

some slight differences were observed in position of nerve ring (slightly posterior to

the base of median bulb vs just below the base of median bulb), shape of female tail

(only long filiform vs variable: short conoid to long filiform). shape of gubernaculum

(slightly pointed proximal end vs blunt). greater a value in males (31 — 39 vs 19.7-

28.4) and in position of phasmids from anus in males (20-24 pm vs 17— 19 pm).

113 Table 11: Measurements (in µm) of Demaniella keirakensis. Mean and S.D. given in parenthesis.

Characters Females (n = 9) Males (n = 10) L 760-999 (896.9 ± 83.9) 698 - 936 (847.5 ± 72.0) a 31.0 - 40.8 (36.2 ± 2.9) 30.2 - 39.0 (35.97 ± 2.3) b 5.4-6.1(5.7±0.2) 4.98-6.5(5.5±0.4)

c 3.4 - 4.5 (4.1 ± 0.3) 3.9-5.5 (4.4 t 0.4)

c' 13.5-21.4(16.6,12.6) 8.1-11.3(9.8±0.97)

V 41.5-48.3(45.1±2.5)

T 43.5-54.5 (49.8 ± 3.9) Maximum body width 20-32 (25.0 ± 3.5) 19-26 (23.6 ± 1.9) Lip width 5-6(5.4±0.5) 4-6(5±0.45)

Length of stoma 10-13(11.1±0.99) 9-12(10.3±0.9)

Pharynx 133-182(157.2±16.7) 135-167(155.3±11.3) Excretory pore from anterior end 123 - 162 (137.9 ± 12.0) 113- 142 (129.5 ± 10.7) Hemizonid from anterior end 118- 155 (131.7 ± 11.2) 107-150(125.8 ± 14.6) Nerve ring from anterior end 110-140(123.3±9.96) 111-130(119.7±7.8) Median bulb (length) 30-40 (33.9 ± 3.1) 30-35 (32.0 ± 1.8) Median bulb (width) 11 - 16 (12.9 ± 1.5) 10- 13 (12.1 ± 0.9) Anterior gonad 108-205 (146.0 ± 33.99) Posterior gonad 107 - 160 (142.2 ± 14.6)

VBD 19-34(26.0±3.7)

Vulva 315-460 (405.1 ± 47.4) Vulva - anus distance 230 - 310 (269.6 ± 30.7) Rectum 16-22(18.8± 1.9) 19-22(20.6±0.9) Tail 190-260(222.2±19.9) 170-205(193.5± 13.6)

ABD 11-17(13.7±2.3) 16-22(19.9±1.6) Phasmids from anus/cloaca 22 - 25 (23.4 ± 0.8) 20 - 24 (21.2 ± 1.4) Testis 315-500(424.5±64.4)

Spicules along axis 29 - 36 (32.0 ± 2.2) Spicules along cord 25 - 30 (27.2 ± 1.6) Gubernaculum 19-23(21.0±1.1)

114

B C

:1 !I1

Fig. 21. Deinaizidllu kezrukens,s. A. Entire temale. B. Entire male, C. Pharyngeal region. D. Female reproductive tract (Anterior), E. Female posterior region. F. Male posterior region.

115 • g C p

'tk .:

11 ~

•

~ .

fri;

1 ~G,,f+. ~ • I

Fig. 22. Dwnaurie'ila kclrake,zsi.s. A. Anterior pharynx. B-I). Anterior region showing stoma & spine-like processes on lips. E. Median bulb, F. Female reproductive tract (Anterior branch). arrow showing bibbed uterus, G. Spermatheca filled %%ith sperms. H. Vulval region. I. Female posterior region , J. K. Male posterior region shoeing spicules & gubernaculum, (Scale bars = 2Opm) Family Neodiplogastridac (Paramonov, 1952) Andrassy, 1984

Diagnosis: Cheilostom divisible into longitudinal furrows in the form of plates or

rods. Cheilostom and gymnostom mostly evenly long. Stegostomal wall asymmetrical, dorsal wall with large, claw-like, movable tooth, right subventral swelling mostly with plate of two or more pointed saw-like teeth or seldom with very small tooth or else entirely unarmed. The posterior section of stoma formed by spacious often prolonged stegostom. Female reproductive system paired. Bursa if present rudimentary. Spicules free. Tail length variable.

Type subfamily: Neodiplogastrinae Paramonov, 1952

Other subfamily: Glauxinematinae Andrassy, 1984

Subfamily Neodiplogastrinae Paramonov, 1952

Diagnosis: Stoma either uniform or divisible into two sections, anterior section wider comprising of cheilostom and gymnostom. Posterior section narrow, often tubular comprising of stegostom, which houses the tooth, teeth or denticles. Cheilostom divisible into rods or plates. Dorsal stegostornal wall with a large movable claw-like tooth and right subventral wall with 1-2 saw-like teeth or small teeth. Spicules free.

T% pe genus: A'eodiplogaster Cobb, 1924

Other genera: Fictor Paramonov, 1952

Koerneria Meyl, 1961

Micolet:kya Weingartner, 1955

Mononchoides Rahm, 1928

Oigolaimella Paramonov, 1952

Prislionchus Kries. 1932

117 j,J Genus Fictor Paramonov, 1952

Diagnosis: Body 0.5 — 2.4 mm long. Cuticle longitudinally striated and punctated.

Labial papillae mostly setae-like. Amphidial openings small, on lateral lips or large,

oval at the base of stoma. Stoma spacious, as long as wide, with a single chamber.

Cheilostom divisible into cuticularized rods. Stegostom with asymmetrical walls,

dorsal and right subventral each with a large, claw-like tooth, left subventral with fine

saw-like plate. Female reproductive system paired. Bursa exceptionally present, 9 —

10 pairs of caudal papillae, with 3 pairs precloacal. Spicules free, relatively short. Tail

in both sexes long, filiform. pointed at tip.

Type species: F. voraa (Goode', 1929) Paramonov, 1952

Fictor suptilis sp. n. (Fig. 23, 24)

Measurements: In Table 12.

Descriptions

Females: Body 0.7 — 0.99 mm long, almost straight upon fixation. Cuticle with longitudinal ridges and transverse striations. Amphidial apertures oval, just below the base of stoma. Lip region continuous with body contour with six fused lips, each lip with a setose papilla. Cheilostom wide with strongly cuticularized chielorhabdions.

Cheilorhabdial flaps extending slightly beyond labial contour, 14 18 in number.

Gymnostom anisotopic, dorsal wall shorter than the subventrals with wart-like denticles. Stegostom anisomorphic. Dorsal wall with a claw-like tooth, right subventral wall with a claw-like tooth and left subventral wall with a serrated plate.

Pharynx about 14 — 19 % of total body length. Procorpus muscular, median bulb ovoid. Isthmus short expanding posteriorly to a basal bulb. Basal bulb with prominent gland nuclei. Nerve ring encircling the isthmus in middle or anterior to middle, 68 —

118 78 % of total pharyngeal length. Hemizonid slightly posterior to the nerve ring, 72 —

81 % of the total pharynx length. Excretory pore posterior to hemizonid, 74 —84 °/ of

total pharyngeal length. Cardia distinct. Intestinal lumen wide.

Reproductive system amphidelphic, anterior branch on right and posterior on

left side of intestine. Ovaries reversed, flexures may or may not overlap each other,

Oocytes arranged in two or several rows. Oviduct narrow. Spermatheca large,

separated from the uterus by a constriction, containing sperms. Uterus divisible in a

short muscular and long glandular regions. Vagina muscular, tubular, one-fourth of

the body width long. Vulva) opening circular. Vulva-anus distance 5.6 — 9.0 vulva)

body diams long. Rectum 0.9 to 1.3 anal body diam. long. Phasmids distinct, 1.5 to 2 anal body diams posterior to anus. Tail long, filiform, 1.5 to 2.5 times vulva-anus distance.

Males: Body ventrally curved in posterior region. Anterior end similar to that of females except for four cephalic papillae present. Testis ventral, reflexed in proliferation zone. Flexure on right subventral side. Spicules long, thin, ventrally arcuate, setose with a very fine tip, 1.6 to 2.9 anal body diams long. Gubernaculum slender with an extension at the proximal end and a sleeve surrounding the spicules at the distal end, 54 — 65 % of spicules length. A cuticular depression present on anterior cloacal lip. Genital papillae setose, eight pairs; two pairs precloacal and six postcloacal. Genital papillae formula: vi, v3d/ v4, ad, phasmids, (v5, v6. v7), pd.

Genital papilla vi within the range of spicules when retracted. GP v2 absent. GP pd far posterior to the (v5, v6, v7) group, in the filiform part of the tail. Tail in two parts; anterior short, conoid part and posterior long filamentous part.

Type habitat and locality: Farmyard manure collected from the District Baramulla,

Kashmir, 1&K, India.

119 Type specimens

Holotype: Female on slide Fictor suptilis sp. n./7 deposited in the nematode collection

of Department of Zoology, Aligarh Muslim University, Aligarh.

Paratypes: Females and males on slides Fictor suprilis sp. n./1-6 deposited in the

nematode collection of Department of Zoology, Aligarh Muslim University, Aligarh.

Diagnosis and relationship

The new species Fictor sup/ills sp. n, is characterized by a small body, wide

lip region, dorsal and right subventral wall of stegostom with a claw-like tooth, left

subventral wall provided with serrated plate, presence of cuticular infolding anterior

to cloaca, long, ventrally arcuate setose spicules, a slender gubernaculum with an extension proximally and a sleeve distally sleeve and eight pairs of setose genital

papillae.

The new species resembles Fictor selosus Mahamood et al., 2006b in having same body length, value of b in both the sexes, value of a in males, pharyngeal length, female reproductive system, presence of cuticular depression on anterior cloacal lip, general shape and length of Spicutes and in number and arrangement of genital papillae. However, the new species can be differentiated from F. setn.su.s in having greater a value in females (28.9 — 42.5 vs 20.8 — 26.8), smaller c value (2.2 — 2.7 vs

3.5 — 5,0 in females and 2.2 — 2,6 vs 3.4 — 3,8 in males), greater c' value (21-4 — 28.6 vs 8.4 — 14.0 in females and 12.8-22.9 vs 8.0 — 11.2 in males), smaller value of V

(35.3 — 38.95 v.r 41.6-51.9), wider lip region (12 — I5 µm ¢c 5 — 9 pm), longer stoma

(10 — 15 pm vs 4-6 pin in females and 8-11 pm vs 6 pm in males), longer tail in both the sexes (300-400 pm vs 107-190 µm in females and 243-320 µm vs 141-

175 pm in males), in the shape and length of gubernaculum (with a extension at the proximal end, 19 — 24 pm in length vs without extension, 15 — 16 µm long), in the shape of labial papillae (short, papillose vs setae-like) and in shape and position of

120 amphidial apertures (oval, below the base of stegostom vs circular pore-like, at the level of stegostom).

Etymology: The species is named after the slender spicules.

121 Table 12: Measurements (in µm) of Fictor suptilis sp. n. Mean and S.D. given in parenthesis.

Characters Holotype Females (n = 10) Males (n = 10) L 828 703-991(869.1±87.3) 592-755(655.1±46.8) a 41.40 28.94-42.5 (37.1 t 4.94) 28.2-35.95 (32.2 ± 2.4) b 6.73 5.4-7.3(6.5±0.6) 4.8-5.9(5.4±0.3)

c 2.51 2.2-2.7(2.5±0.2) 2.2-2.6(2.4±0.1) c' 27.50 21.4-28.6 (25.3 ± 2.1) 12.8-22.9 (16.7 ± 2.8) V 37.56 35.3 - 38.95 (37.4 ± 0.97) T 23.1 -33.1 (28.6±2.9) Maximum body width 20 20-34 (23.9 t 4.7) 18 - 22 (20.4 t 1.1) Lip width 12 12-15(12.8±0.98) 8-13(11.4 t 1.3)

Length of stoma 10 10- 15 (11.0 f 1.6) 8- 11 (9.6 t 0.8) Pharynx 123 123 - 158 (133.6 f 8.9) 106 - 130 (122.4 ± 7.0) Excretory pore from anterior end 103 97 - 120 (104.3 f 6.1) 83 - 109 (97.0 ± 8.5) Hem izonid from anterior end 99 94 - 117 (100.4 t 6.7) 79 - 103 (92.9 ± 8.2) Nerve ring from anterior end 90 90 - 115 (95.1 ± 8.0) 75-94(85.1±5.7) Median bulb (length) 19 19-30(21.7±3.4) 17-25(20.5±1.9) Pharynx base to gonad 117 90-120(I01.9±9.1) 48-100(73.2±15.2) Anterior gonad 71 40 - 131 (90.3 t 29.96) Posterior gonad 87 40-125(86.4±25.5)

VBD 23 22-36(25.9±4.7) Vulva 311 266 - 386 (325.8 ± 37.5) Vulva - anus distance 187 135-233 (192.1 ± 30.9) Rectum 15 13-20(15.4± 1.9) 14-20(16.6±2.2) Tail 330 300-400 (351.1 ± 27.4) 243 - 320 (272.3 ± 24.6) ABD 12 12-15(13.9±0.8) 14-23(16.7±2.7) Phasmids from anus/cloaca 19 19-30 (22.5 ± 3.1) 18-25 (21.4 ± 2.2) Testis 160-250 (187.2 ± 26.1) Spicules along axis 35 - 40 (37.0 ± 1.7) Spicules along cord 31 - 35 (32.8 ± 1.1) Gubernaculum 19 - 24 (21.2 ± 1.8)

122 A ~rB ç a c Ad F ur'

L ~

C

Fig. 23. Fictor .suptilis sp. n. A. Entire female, B. Entire male. C. Anterior region shop%ing amphids dorsoventrally, 1). Anterior region (lateral). V. Pharyngeal region, F. Female reproductive tract (Anterior), G. Female posterior region. I1. Male posterior region (lateral). I . Male posterior region (dorsoventral).

123 1.

A B C D E

~.. I G

•s

ii T d

F

• K 1'

~•~ :i L i I

Fig . _' 1 l r •r ,cru/,li/i p. i :\..\ntei iur rc_iun'showing stoma. U. Anterior region showing dorsal claw-like tooth, C. Anterior region showing dorsal tooth & right sub~entral tooth. D. Anterior region showing Cheilorhabdions & denticles. E. Anterior region showing amphids dorsoventrally, F. region showing serrated plate. G. Pharyngeal region. H. Cuticle shoeing longitudinal ridges, I. Female reproductive tract (anterior branch). J. K. Male posterior region showing spicules & gubernaculurn, (Scale bars — 20µm). Fictor longicauda sp. n. (Fig. 25, 26)

Measurements: In Table 13.

Descriptions

Females: Body almost straight upon fixation. Cuticle with prominent longitudinal

ridges and transverse striations. Amphidial openings, prominent, large, oval, slightly

posterior to the base of stoma. Lips fused. Lip region continuous with body contour

with setose labial papillae. Cheilostom wide, cuticularized, cheilorhabdions rod-like

approximately 18-20 in number, directed inwards apically. Cheilorhabdial flaps bifid

extending beyond labial contour. Gymnostomal walls strongly cuticularized, provided

with denticles. Stegostom anisomorphic, dorsal and right subventral wall with a claw-

like tooth; left subventral wall with serrated margin divisible into left and right plates.

Pharynx 13 — 16 % of total body length. Procorpus muscular, almost of uniform

diameter throughout. Median bulb oblong with strong valve plates. Isthmus short,

gradually expanding to form basal bulb. Anterior pharynx 65 — 71 % of total

pharyngeal length. Nerve ring encircling isthmus just below the median bulb.

Hem izonid posterior to nerve ring, 72 — 80 % of the total pharyngeal length. Excretory

pore 75.7 - 85 % of the total pharyngeal length. Cardia well developed. Intestine with

wide lumen. Intestinal cells large.

Reproductive system amphidelphic. anterior branch on right and posterior arm

on left side of intestine. Ovaries reversed, flexures may extend beyond vulva. Oocytes arranged in two or more rows. Oviduct long and narrow, gradually expanding to form a spermatheca. Spermatheca large, not set off from the uterus, usually with sperms.

Uterus with long glandular and short muscular parts. Vagina muscular, tubular, one- fourth of the body width long. Vulval opening circular. Rectum 1.2 — 1.3 anal body

125 diam. long. Phasmidial apertures prominent, approximately one anal body diam.

posterior to anus. Tail long, filiform 2,5 —2.8 times vulva-anus distance.

Males: Body curved in posterior region. Anterior end similar to that of females except

four cephalic papillae present. Testis ventral, reflexed in proliferation zone. Flexure

on right side. Spicules slender, ventrally arcuate. 1.7-2.3 times anal body diarn. long.

Gubernaculum simple, broad gradually tapering distally, with a short hook-like

extension proximally and a sleeve distally, 46 — 55 % of spicules length. Genital

papillae nine pairs, setose; 2 pairs precloacal and 7 postcloacal. Genital papillae

formula: vl, v3d / v2, v4, ad, phasmids. (v5, v6, v7), pd. GP vl within the range of

spicules when retracted. OP v2 just below the cloaca. GP pd far posterior to v5-7

group in the filiform part of tail. GP v7 longer than v5 and v6. Tail long filiform.

Type habitat and iocalliy: Decayed banana rhizome collected from Haldwani,

Uttrakhand, India.

Type .specimens

Holotype: Female on slide Ficror longicauda sp. n./7 deposited in the nematode

collection of Department of Zoology, Aligarh Muslim University, Aligarh.

Pararvpes: Ten females and ten males on slides Ficror longicauda sp. n./1-6 deposited

in the nematode collection of Department of Zoology, Aligarh Muslim University,

Aligarh.

Diagnosis and relationship

Fictor longicauda sp. n. is characterised by a medium sized body, presence of several warts on the gymnostomal walls, left suhventral stegostomal wall with serrated margin divisible into two plates. slender ventrally arcuate spicules, gubemaculum broad proximally and tapering distally with a proximal hook-like extension and a distal sleeve, and nine pairs of genital papillae, GP v2 just below the cloaca and pd far posterior to GP v7 in the filiform part of the tail.

126 The new species resembles Fictor denticulatus Mahamood et al., 2006b in having same body morphology and morphometrics. However the new species differs from F. denticulatus in having a longer tail (c = 1.98 - 2.1 vs 2.4 - 2.9 in females and

2.0 - 2.5 vs 2.7 - 3.6 in males), in anterior position of value (V = 31.2 - 33. 5 vs 36.2

- 40.1), greater lip width (15 - 18 pm vs 9 - 10 µm), position of nerve ring (at the base of median bulb vs almost at the middle of isthmus), greater ABD (16- 19 µm vs

13 - 14 µm), cuticular depression at upper cloacal lip (absent vs present) in position of phasmids from anal opening (27 - 32 µm vs 15 - 17 µm), shape of gubernaculum

(simple, gradually tapering distally with a proximal hook-like extension and a distal sleeve vs broad wedge shaped abruptly tapering distally without proximal extention and distal sleeve), and in number and arrangement of genital papillae (nine pairs, GP v2 present at the lower cloacal lip vs eight pairs, GP v2 absent).

Etymology: The name of the species is derived from the long tail.

127 Table 13: Measurements (in µm) of Fictor longicauda sp. n. Mean and S.D. given in parenthesis.

Characters Holotype Females (n = 10) Males (n = 10) L 850 785 - 955 (884.5 t 56.2) 579-926 (718.1 ± 86.7) a 38.64 29.6-38.6(33.6±2.9) 32.2-44.1 (35.8±3.7)

b 6.59 6.2-7.5(6.8±0.4) 5.4-7.98(6.5±0.7)

c 1.98 1.98-2.1(2.0±0.1) 2.0-2.5(2.2±0.1)

co 25.29 22.2-27.8 (24.8 ±1.7) 14.4-21.7 (18.4±2.2) V 31.18 31.2-33.5 (32.5 ± 0.8) T 25.1 - 39.4 (33.2 ± 4.2) Maximum body width 22 22 - 30 (26.5 ± 2.3) 17 - 23 (20.1 f 1.6) Lip width 17 15-18(16.7±0.8) 11-15(13.1±1.2)

Length of stoma 12 10-12(11.2±0.8) 7-9(8.2±0.6)

Pharynx 129 115- 139 (130.6 t 7.2) 97 - 120 (110.0 t 6.3) Excretory pore from anterior end 101 95 - 116 (106.9 f 6.9) 80 - 103 (90.4 t 7.9) Hem izonid from anterior end 97 90 - 111 (101.5 f 6.8) 75 - 97 (86.7 t 7.9) Nerve ring from anterior end 94 87 - 100 (95.1 t 3.9) 71- 86 (77.7 t 5.7) Median bulb (length) 24 18-26(24.2 ± 2.2) 15-21(18.1 t 2.1)

Median bulb (width) 17 13-20(17.6 ± 2.1) 11 - 15 (13.3 t 1.2) Pharynx base to gonad 75 59-101(78.0±14.0) 30-60(45.1 t 10.4) Anterior gonad 61 60-90(78.8± 11.3)

Posterior gonad 71 40 - 125 (77.5 t 21.0) VBD 22 22-30 (26.4±2.3) Vulva 265 250 - 311 (287.4 ± 22.0) Vulva-anus distance 155 148-177(164.1 ± 10.4) Rectum 18 16 - 20 (17.6 ± 1.3) 19 - 25 (22.2 t 1.9) Tail 430 380 -475 (433.0 ± 27.3) 260- 400 (322.5 ± 37.0) ABD 17 16-19(17.5±1.0) 15-20(17.6± 1.5) Phasmids from anus/cloaca 29 27-32 (29.7 ± 2.1) 24 -33(27.4 ± 2.7) Testis 165 - 365 (240.5 ± 54.1)

Spicules along axis 32 - 37 (35.0 ± 1.9) Spicules along•cord 29 - 34 (3).6 ± 1.6) Gubemaculum 15-18(17.0±1.0)

128 •

A,B1 --150µm

--4 20 µm

D

Fig. 25. Fictor luntgicai thi sp. H. A. Fntire temale, I. Entire male. C. Anterior end d rrsoventral ). 1). Phary n:;eal region. F. Female reproductive tract (Anterior). F. Female posterior region, G. Male posterior region.

129

ii t \u\ f m E

fi

f I$• I \/

1". 2b. !•recur Ju#zgie auta sp. n. A. Pharyngeal region, 13. Anterior region sho%%ing Cheilorhabdions & cheilorhabdial tlaps, C. Anterior region showing dorsal claw -like tooth & right subventral tooth, P. Anterior region sho«ime cheilorhabdions & , mnostomal wall with denticles. F. Cuticle showing longitudinal ridges. F. Female reproductive tract (anterior branch), (I. Female posterior region, H. I. Male posterior region showing spicules & gubernaculum, J. Male posterior region showing genital papillae, (Scale bars ?0µm). Genus Koerneria Meyl, 1961

Diagnosis: Body length 0.3-1.7 mm. Cuticle with fine transverse and longitudinal

striations. Labial papillae minute. Stoma small, almost as long as wide. Cheilostom

and gymnostorn short, cuticularized. Stegostomal walls provided with a claw-like

tooth each on the dorsal and right subventral walls, left subventral with a ridge

provided with small denticles. A pair apodemes, one each on the subventral walls,

present. Female reproductive system paired or unpaired. Spicules arcuate, slender.

Tail long, Rliform in both sexes.

Type species: Koerneria goffarti (KBrner, 1954) Meyl, 1961 — Diplogaster

(Eudiplogaster) goffarli Kbrncr, 1954

Koerneria friitaudata (Khera, 1970) Andrassy, 1984 (Fig. 27, 28)

Measurements: In Table 14.

Descriptions

Females: Body almost straight upon fixation, gradually tapering posteriorly. Cuticle

finely transversely striated, longitudinal ridges present. Punctuations present, arranged

both in transverse and longitudinal rows. Amphidial apertures elliptical, at base of

stoma. Lip region continuous with body contour with six fused lips. Each lip with a

setose labial papilla. Stoma wide and shallow. Cheilostom cuticularised with six plate-like cheilorhabdions. Each plate extending beyond labial contour. Gymnostomal

walls more or less similar, arching inwards. Dorsal stegostomal wall with a big claw- like tooth, right subventral wall with a similar cuticularized tooth and left subventral wall with a serrated margin. Posterior end of stegostom with prominent apodemes on subventral walls. Procorpus muscular, median bulb ovoid with strong valve plates.

Isthmus short, expanding to a basal bulb. Anterior pharynx muscular, 65 — 71 % or

131 pharyngeal length. Nerve ring encircling isthmus, 72 - 81 % of total pharyngeal

length. Hemizonid slightly posterior to nerve ring, 66 - 85 % of total pharyngeal

length. Excretory pore further posterior to hemizonid, 83 % of total pharyngeal

length. Cardia present. Intestine with wide lumen.

Reproductive system amphideiphic. anterior branch on right side and posterior

on left side of intestine. Ovaries reversed, oocytes arranged in one or more rows.

Oviduct short, gradually expanding to form a spermatheca. Spermetheca round or oval in shape, separated from uterus by a constriction. Uterus divisible into and thick glandular and a short muscular part. Vagina muscular, about one-third of vul~al body diam. long. Vulval opening circular. Vulva-anus distance about 7.6 - 10 times vulval body diam. long. Rectum I - 1.4 times anal body diam. long. Phasmids prominent, one to two anal body diam. posterior to anus. Tail filiform, whip-like, about 21 -- 31 times anal body diam. long.

Males: Body usually smaller than females, curved in posterior region. Anterior region similar to that of females except for four cephalic setae. Spicules slender, curved, about 1.4 - 1.6 times anal body diam. long. Gubernaculum boat-shaped, 55 - 69 % of spicules length, \%ith a distal sleeve. Genital papillae well developed. setose, nine pairs: three pairs precloacal, six pairs postcloacal. Genital papillae formula: vl, v2, v3d - v4, ad. phasmid, (v5, v6, v7), pd. vI beyond the range of spicules when retracted. v2 and v3d almost at the same level. Phasmids posterior to v4, v7 at a slight distance from v5, 6 and pd well posterior to the v5-7 group. Tail divisible into two parts, a short conoid part and a long filiform part.

Habitat and localitt: Farmyard manure collected from Department of Zoology, AMU,

Aligarh, India.

132 Voucher specimens

Ten females and ten males on slides Koerneria filicaudata/l -6 deposited in the

nematode collection of Department of Zoology, Aligarh Muslim University, Aligarh.

Remarks

The measurements and descriptions of our specimens agree well with the original specimens given by Khera, 1970. However, differences were found in having slightly greater a value (36.9 — 45.6 vs 34 — 36), greater b value (6.8 — 8.2 vs 5.7 —

6.2) and slightly longer stoma (8 — 10 pm vs 7.5 — 8.5 µm). The measurements and descriptions of our specimens also conform well with that given by Mahamood and

Ahmad (2010). However, slight differences were found in having greater a value

(36.9 — 45.6 vs 28.8 — 35.9), greater b value (6.8 — 8.2 vs 5.6 — 6.6), shorter stoma (8 —

10 µm vs 11 — 13 µm) and shorter reflexed part of ovary.

133 Table 14: Measurements (in µm) of Koerneria frlicaudata. Mean and S.D. given in parenthesis.

Characters Females (n = 10) Males (n =10) L 645-875(801.2=69.4) 610- 729(668.81 31.6) a 36.9- 45.6 (40.3 t 2.7) 38.3-44.9(41.6±1.9) b 6.8-8.2(7.5±0.4) 5.9-6.8(6.4 ± 0.3) c 2.3-29(2.610.2) 2.7-3.2(2.910.2) c' 20.8-30.8(26.1+2.9) 13.1-17.9(15.3+1.6) V 34.1 -40.8(36.8±2.1) T 28.9-41.5(35.9±3.9) Maximum body width 17-23 (19.9 t 1.6) I5-17(16.1 t 0.8) Lip width 10-12(11.5±0.7) 9-10(9.9 t 0.3) Length of stoma 8-10(9.5±0.7) 6-8(7.0±0.5) Pharynx 92-117(107.1 16.7) 96-110(104.5±4.9) Hemizonid from anterior end 68-99(85.7 + 13.0) 77-90(82.3 ±5.6) Nerve ring from anterior end 66-87(81.6 ± 73) 77 - 94 (82.0 ± 6.2) Median bulb(length) 15-19(16.9±1.!) 14-17(15.011.0) Median bulb (width) 11-14 (12.6 t 1.0) 9-12(10.5+0.8) Pharynx base to gonad 82- 128 (]05.5 f 15.2) 54-130(90.8±24.3) Anterior gonad 60- 120 (84.3 t 16.8) Posterior gonad 50-91 (80.8211.6) VIM) I8 -26(21.912.4) Vulva 220-357 (295.5 t 35.1) Vulva-anus distance 155-218(193.7± 17.3) Rectum 13-16(14.6±1.6) 15-16(15.5±0.5) Tail 270-370(312.0±30.3) 210- 260 (233.0 f 16.8) ABD 10-13(12.0+0.9) 14-17(15.3±0.9) Testis 195 - 292 (240.5 z 29.8) Spicules along axis 21 - 23 (22.2 f 0.8) Spicules along cord 19 -20 (19.510.5) Gubemaculum 12-16(14.4±1.0)

134 LJ

i ! •~, ~n

Hcl U

C

Fig. 27. Koerneria ,hlicauduta. A. Entire female, B. Entire male, C. Phar%negeal region. D. Female reproductive tract (entire), E. Female posterior region, F. Male posterior region.

135 A B C E

II I R `

}

H ` r te I

• F

Fig. 28. k'uerneriu /ilic•uuduru. A. Pharyngeal region. 13, C. Anterior region showing stoma, D. Anterior region showing Cheilorhabdions & serrated ridge, E. Female reproductive tract (anterior branch), F. Vulval region, G. Female posterior region. It. I. Male posterior region sho %ing spicales & gubernaculuni. (Scale bars 20µm). Genus Mononc/aides Rahm, 1928

Diagnosis: Body length 0.6 — 3.4 mm. Cuticle with transverse striations and

longitudinal ridges. Labial papillae setae-like. Stoma distinctly divisible into two

parts. The anterior section comprising of cheilostom and gymnostom. The posterior,

narrow, tubular part comprises of stegostom. Cheilostomal walls always divisible into finely pointed rods (12 — 18 in number). Dorsal stegostomal wall with a large movable claw-like tooth, left subventral with immovable transverse plate and right with saw-like teeth. Female gonads paired. A true but week bursa frequently present.

Spicules free, moderately slender. Gubemaculum proximally always incised. Genital papillae 9 or 10 pairs of which 3 are precloacal. Tail in both sexes long, filiform.

Type species: Al. longicauda Rahm, 1928

Mononchaides tenuispicula sp. H. (Fig. 29, 30)

Measurements: In Table 15.

Descriptions

Females. Body long, straight to slightly arcuate ventrally in fixed material. Cuticle with conspicuous longitudinal ridges, 16 — 18 in number. Lip region continuous with body contour, lips fused, each with a small papilla. Amphidial apertures large, oval. located at base of gymnostom. Stoma longer than wide. Chiclostom wide, heavily culicularized. Cheilorhabdions rod-like, bifurcated anteriorly. Gymnostom anisotopic with subventral walls longer than dorsal. Stegostom anisotopic and anisomorphic; dorsal wall bearing a large claw— like dorsal tooth, right subventral wall with a pyramidal tooth and Icft subventral with a denticulate ridge. Posterior part of stegostom broad and tubular. Pharynx 12 to IS % of total body length, divided into muscular procerpus expanding posteriorly into oval, muscular median bulb with prominent valve plates. Isthmus short, gradually expanding into an elongated,

137 glandular basal bulb. Nerve ring encircling isthmus in the anterior half, at 65 to 83%

of pharyngeal length from the anterior end. Hemizonid slightly posterior to nerve ring.

Excretory pore posterior to hemizonid, at 76 to 90 % of pharynx. Excretory duct

anteriorly directed. Cardia distinct.

Reproductive system amphidelphic, both branches almost equally developed.

Anterior branch on right and posterior branch on left side of intestine. Ovaries

reversed with the terminus of each reaching upto vulva or beyond. Oocytes arranged

in two or more rows in germinal zone. Oviduct short and narrow. Spermatheca

continuous with uterus, not offset, containing sperms usually. Uterus with a large,

prominent glandular part and a smaller and narrow muscular part. Vagina muscular,

opening circular, pore like. A pair of dumb-bell shaped pouches present at level of

uterus. Vulva-anus distance about 12 to 14 anal body diam. long. Rectum 0.95 to 1.32

anal body diam. long. Phasmids prominent, located 0.8 to 1.1 anal body diam.

posterior to anus. Tail long. filiform about 1.3 to 1.8 vulva— anus distance long.

Males: Body smaller, anterior end similar to that of females except for four additional cephalic setae. Testis single, reflexed. Spicules arcuate, proximal end with a rounded head, 1.72 to 1.84 anal body diam. long. Gubernaculum narrow, keel-shaped with a roughly pointed proximal end and a fine distal tip provided with a sleeve. Nine pairs of genital papillae: three pairs precloacal and six pairs postcloacal. Genital papillae formula: vl, v3d, v2 / v4, ad, phasmids, (v5, v6, v7), pd. Genital papilla (GP) v2 located at the level of upper cloacal lip. GP ad located close to phasmids. Tail divisible into a short conoid and a long filamentous part.

Type habitat and loca/in': Farmyard manure collected from Umar Colony. Baramulla.

Kashmir.

138 Type specimens

Holotype: Female on slide Mononchoides tenuispicula sp. n./8 deposited in the

nematode collection of Department of Zoology, Aligarh Muslim University, Aligarh.

Paratypes: Ten females and nine males on slides Mononchoides tenuispicula sp. n./l-

7 deposited in the nematode collection of Department of Zoology, Aligarh Muslim

University, Aligarh.

Diagonosis and relationships

The new species is characterized by a wide tubular part of stegostom, anteriorly directed excretory duct, narrow, arcuate spicules provided proximally with

rounded head and a narrow, keel-like gubernaculum with roughly pointed proximal end, narrowing distally to a fine tip provided with a sleeve.

The new species resembles ttfononchoides gaugleri Siddiq i, Bilgrami and

Tabassum, 2004 in having same morphometric values, body length and in number and arrangement of genital papillae. However, the new species can be differentiated from

Mononchoides gaugleri in the shape of gubernaculum (narrow keel-shaped with a fine distal sleeve vs distal fourth surrounding the spicules), in the position of phasmids with respect to the anal opening (15 — 20 gm vs 22 — 25 µm), excretory duct

(anteriorly directed vs posteriorly directed) and in vaginal glands (absent vs present; four pairs).

The new species further resembles Mononchoides adjunctus Massey, 1966 in general morphometrics, body length, structure of stoma. However, the new species can be differentiated from Al adjucntus in position of nerve ring from anterior end

(encircling anterior half of isthmus vs middle of isthmus), shorter tail (c = 2.6 — 2.9 vs

3.2). in length and shape of spicules (31 — 36 }un. almost uniformly arcuate ventrally vs 43.3 µm, strongly curved in the anterior half), structure of gubernaculum (keel-like with pointed proximal end; distal sleeve present vs almost rounded proximal end;

139 distal sleeve absent) and in number of genital papillae (nine; three preclancal, six

post-cloacal vs seven; two pre-cloacal, five post-cloacal).

The new species resembles Mononchoides chungi Googrich, Hechler and

Taylor, 1968 in general morphology and morphometriy values and in body length.

However, the new species can be differentiated from Mononchoides changi in the lip region (continuous vs offset), in the position of excretory pore (112— 129 µm vs 143-

150 pin), in the length and shape of spicules (31 — 36 pm, uniformly arcuate vs 48 —

49 µm, proximal part slightly dorsally inclined) and in shape of gubermculum (with a distal sleeve, not constricted vs deeply constricted, without sleeve).

The new species also resembles Mononchoides bollingeri Googrich, Hechler and Taylor, 1968 in having same morphornetric values, pharynx length and position of nerve ring and excretory pore. However, the new species can be differentiated from

Mononchoides bollingeri in body length (0.95 — 1.11 mm vs 1.64 — 3.43 mm in females and 0.64 — 0.83 min vs 1.00 — 2.40 mm in males), in the length of tubular part of stegostom (short vs long), in number and arrangement of genital papillae (nine; 3 pairs pre-cloacal, 6 post-cloacal vs ten pairs; 3 pairs pre-cloacal, 7 post-cloacal) and in structure ofgubemaculum (with a distal sleeve vs without a distal sleeve).

The new species further resembles Mononchoides gracilis Dassonville and

Heyns, 1984 in having same body length, in c' value in both the sexes, in b value in males, in tail length in both the sexes and in number of genital papillae. However, the new species can be differentiated from M gracilis in having less slender body (a

27.74-31.69 vv 34 -45), in slightly anterior position of vulva (V 34.31= — 36.96 vs

38 — 425), in structure of gubernaculum (keel-like, with distal sleeve vs sickle shaped, without a sleeve) and in the position of nerve ring (encircling anterior half of isthmus vs middle of isthmus).

Etymology- The species is named after the thin spicules.

140 Table 15: Measurements (in µm) of Alononchoides tenuispicula sp. n. Mean and S.D. given in parenthesis.

Characters Holotype Females (n = 10) Males (n = 9) L 936 958-1117(1042±56.5) 640-831(744±54.6)

a 29.3 27.7-31.7(30.1±1.1) 25.6-34.6(29.4±2.5)

b 6.9 6.7-7.9(7.2±0.4) 5.8-8.1(6.5±0.6) c 2.7 2.4 - 2.9 (2.5 ± 0.2) 2.6-2.9 (2.7 ± 0.1) c' 17.5 15.7-22.6 (20.3 ± 1.8) 12.8-14.5(13.7±0.5) V 37.2 34.3 - 36.96 (35.97 ± 0.8)

T 33.7-43.2 (38.98 f 2.8) Maximum body width 32 31 - 37 (34.5 ± 2) 24 - 27 (25 ± 1) Lip width 13 14-15(14.5±0.5) 10-12(10.4±0.7) Length of stoma 16 15-19(17±1.1) 11-14(12.6±1.1) Pharynx 136 138-150(143.9±3.7) 103-126(114.7±6.8)

Excretory pore from anterior end 114 112- 129 (120.8 ± 5.7) 90 - 106 (97.6 f 5.1) }lemizonid from anterior end 110 107- 124 (115.4 ± 5.2) 86 - 97 (90.8 t 4.2) Nerve ring from anterior end 95 92 - 121 (98.9 ± 7.8) 71 - 85 (79.2 t 4.8) Median bulb (length) 24 22-26(24± 1.4) 16-21(18.4±1.6) Anterior gonad 142 130 - 195 (164.1 ± 19.2) Posterior gonad 153 130-176(151.2±13.2) VBD 33 30-38 (35 ± 2.5) Vulva 348 347 - 404 (377.4 ± 17.9) Vulva - anus distance 238 225-280 (253.3 ± 15.4) Rectum 22 19 - 25 (22 ± 1.9) 18 - 25 (20.2 ± 1.9) Tail 350 330-460(411.3±36.1) 245-300(273.3± 15.6) ABD 20 19-21 (20.3}0.9) 18-21 (19.9±0.9) Phasmids from anus/cloaca 18 16-24(19,9± 1.9) 15-20(17.3± 1.8) Testis 230 - 335 (290.3 ± 29.5) Spicules along axis 31-36(34.1 t 1.4) Spicules along cord 28 - 32 (30 t 1.1) Gubemaculum 14-18(16.2±1.0)

141 F

L b>wI a

r7 9E

Fig. 29. JJonondw,dt'.s k'nuispiculu sp. n. A. Entire tniale. B. Entire male, C. D. Anterior end (lateral. F. Anterior end (dorsoventral). F. Pharyngeal region, G. Female reproductive tract (Anterior), H. Female posterior region. I. J. Male posterior region.

142

A B C n

(\E

G

H M K l- i". "(). t/uNmlc'1lulJc'.a lc'p11s1\/ 1icula sp. n. ;1. anterior region doming ,ionma, B. Anterior region shoN%ing denticulate ridge. C. Anterior region showing, amphids dorso entrally. D. Anterior region showing Cheilorhabdions, Ii. Anterior pharynx, F. Median bulb & posterior pharynx, G. Cuticle of anterior region. II. Cuticle showing longitudinal ridges. I. Joining of oviduct & oar. J. Vukal region. K. L. Male posterior region showing spicules &. ;gubernaculum, M. Male posterior region showing genital papillae. (Scale bars - 20µm). Mononchoides latigubernacula sp. n. (Fig. 31, 32)

A1eusurements: In Table 16.

Descriptions

Females: Body almost straight upon fixation. Cuticle with transverse striations and

prominent longitudinal ridges. Lip region round, continuous with body contour. Lips

six fused, each with a small papilla. Amphidial apertures oval slits, located at base of

dorsal tooth. Stoma longer than wide, divisible into anterior wide and posterior

narrow parts. Chielostom wide, cuticularized. Chielorhabdial flaps anteriorly

bifurcated. Gymnostom anisotopic, subventral walls longer than dorsal. Stegostom anisotopic and anisomorphic. Dorsal wall with a claw-like tooth with anteriorly directed tip. Right subventral wall with an anteriorly directed pyramidal tooth and left subventral wall with a serrated plate/denticulate ridge. Posterior part of stegostom tubular. Pharynx about 13 % of total body length. Procorpus muscular, gradually enlarging to form a spheroid median bulb. Median bulb 20 — 25 µm long, lumen strongly cuticularized forming an elongate oval valvular apparatus. Isthmus short. gradually expanding into a glandular basal bulb. Anterior pharynx 61 to 68 % of pharyngeal length. Nerve ring encircling isthmus in the anterior half. Hemizonid posterior to nerve ring at about 81 % of pharynx length from anterior end. Excretor\ pore further posterior to hemizonid. ('ardia well developed. Intestinal lumen wide.

Reproductive system amphidelphic. ovaries reversed with the terminus of each ovary often reaching upto vulva. Anterior branch on right and posterior branch on left side of intestine. Ovary long, oocytes arranged in several rows in germinal zone.

Oviduct narrow, expanding to form an elongate spermatheca. Spermatheca separated from uterus by a constriction. Sperms may be present both in the spermatheca and in the oviduct. Uterus with a narrow, muscular distal and a prominent glandular

144 proximal parts. Uterine eggs oval with smooth walls. Vagina muscular. Vulval

opening circular, pore-like. Vulva-anus distance about 6 to 9 anal body diams long.

Rectum 0.9 to 1.4 anal body diam. long. Phasmids almost one anal body diam.

posterior to anus. Tail long filiform with a fine tip about 1.3 to 1.8 times vulva-anus

distance.

Males: Body smaller than the females, ventrally curved in the posterior region.

Anterior region similar to that of females except for four cephalic setae. Testis single,

reflexed. Spicules long, setose, strongly ventrally curved, 1.7 to 2.0 anal body diam.

long with a proximal rounded head and finely pointed distal tip. Gubernaculum broad,

keel-like with blunt proximal end and a sleeve distally, 40 to 50 % of spicules length.

Genital papillae nine pairs; three pairs precloacal, six pairs postcloacal. Genital

papillae formula: v1, v3d, v2/ v4, ad, phasmids, (v5, v6, v7), pd. Tail divisible in two

parts: a short conoid part and a long filamentous part about 13 to 16 anal body diam.

long.

Type habitat and locality: Floating island Khornapaat, Loktak Lake, Manipur.

Ttipe specimens

Holot►pe: Female on slide Afononchoide's latiguhernacula sp. n./6 deposited in the

nematode collection of Department of Zoology, Aligarh Muslim University, Aligarh.

Paratvpes: Eleven females and ten males on slides Mononchoides latigubernacula sp.

n./1-5 deposited in the nematode collection of Department of Zoology. Aligarh

Muslim University, Aligarh.

Diagonosis and relationships

The new species is characterized by a wide cheilostomal region, short and

wide tubular part of stegostom, strongly curved setose spicules and a broad, keel- shaped gubernaculum with a small distal sleeve.

145 The new species resembles bfononchoides gauger Siddiq i, Bilgrami and

Tabassum, 2004 in body length, tail length, morphometric values, length of spicules

and in number and arrangement of genital papillae. However, the new species can be

differentiated from M. gaugleri in shape of gubernaculum (keel-shaped with a small

distal sleeve vs claw-like with distal fourth forming a tube around the spicules), in

vaginal glands (absent vs four pairs of vaginal glands radially arranged around vagina)

and in the position of phasmids with respect to ad (a few microns posterior to ad vs at

the base of ad).

The new species also resembles Mononchoides changi Goodrich, Hechler and

Taylor, 1968 in body length and morphometric values. However, the new species can

be differentiated from M changi in the position of excretory pore from anterior end

(93 — 131 µm vs 143 — 150 pm), in lip region (continuous with body contour vs

offset). in the length and shape of spicules (40 —45 µm long, setose, uniformly curved

vs 48-49 tm long, curvature not uniform, proximal end slightly dorsally directed), in

the shape of gubemaculura (broad keel-shaped with a distal sleeve vs narrow keel-

like, without a distal sleeve).

The new species further resembles Mononchoides aquaricus Dassonville and

Heyns, 1984 in having same b value in males as well as in females, in female body length, in having same number of genital papillae, in having same size of cheiloslom and gymnostom. However, the new species can be differentiated from Mononchoides aquaricat in the size of stegostom (tubular part longer than wide vs as long as wide), in having longer tail (c = 2.5 — 2.8 vs 3.8 — 4.8 in females and 2.4— 2.7 vs 5.1 — 6.0 in males), c' (19.1 — 26.5 vs 9.0— II. in females and 13.0—I 5.5 vs 5.4 — 6.4 in males), position of vulva (I%= 36-38 vs 43 — 49), in length and shape of spicules (40 - 45

µm long. selose, uniformly curved vs 28.8 — 32.0 pm long, well developed, curvature at the proximal half), shape of geubernaculum ( keel-shaped with a distal sleeve vs

146 stirrup-shaped with a needle-like distal process) and in the arrangement of genital papillae (vi within the range of spicules when retracted vs beyond the range).

Etymology: The species is named after the broad gubernaculum.

147 Table 16: Measurements (in µm) of Mononchoides latigubernacula sp. n. Mean and S.D. given in parenthesis.

Characters Holotype Females (n = 11) Males (n = 10) L 984 916-1361(1047±138) 740-925(846±49) 27.3 - 33.7 (29.98 f a 31.7 25.8-36(30.9±2.8) 1.8)

b 8.56 6.3-9.7(8.0±0.8) 7.2-9.4(8.1±0.7) c 2.5 2.5 - 2.8 (2.6 ± 0.1) 2.4-2.7 (2.5 ± 0.1)

c' 24.4 19.1-26.5(22.9±2.1) 13.0-15.5(14.4±0.8)

V 35.7 35.3-37.9 (36.6±0.97)

T 36.9-43.4 (40.8 ± 2.3) Maximum body width 31 27-46 (34.3 t 6.4) 25 - 32 (28 ± 2.2) Lip width 16 15-17(16±0.7) 11- 13(11.9±0.8) Length of stoma 14 13-18(15±1.2) 10-14(11.7±1.1)

Pharynx 115 112-156(130.3±12.1) 95-120(105.3±8.5) Excretory pore from anterior end 97 93 - 131 (109 ± 10.8) 74 - 100 (88.8 f 7.9) Hemizonid from anterior end 93 89 - 124 (103.5 ± 9.4) 70-95 (84.5 ± 7.4) Nerve ring from anterior end 87 76 - 102 (91 f 7.3) 63 -79(72.7 ± 5.3) Median bulb (length) 23 20-25 (22.1 f 1.8) 15 - 20 (17.7 * 1.6) Anterior gonad 161 100-255 (153.4 ±43.8) Posterior gonad 163 93 - 225 (142.6 f 45.5) VBD 32 27-50 (34.8 t 6.8) Vulva 351 330-481(383.4±47.2)

Vulva-anus distance 243 209-370 (258.8 ± 51.3) Rectum 20 18 - 27 (21.6 ± 2.6) 18 - 26 (22.3 ± 2.8) Tail 390 360 - 510 (405.3 ± 42.4) 300-375 (334 ± 23.3) ABD 16 15-24(17.9±2.97) 20-25 (23.3 ± 1.5) Phasmids from anus/cloaca 16 17-24 (19.6 t 2.3) 15 - 22 (18.9 ± 2.4) Testis 305 - 385 (345 ± 29.2) Spicules along axis 40 - 45 (43.3 ± 1.6) Spicules along cord 35 - 40 (37.1 ± 1.3) Gubemaculum 20 - 22 (20.5 ± 0.7)

148 D

a E20 um

kb ill!!

1-1

I

Fig.. 31. Mono,,dwides lutigubernuculu sp. ii. A. lntire tmaIe. B. Entire male. C- F. Anterior uid. F. Pharyngeal region. (i. Female reproductic tract (Anterior). H. Female posterior region, I. Male posterior region.

149

A B ' C ( s~ G

k E

F I H I. I l:.* , q

!1

Y 1 i rç 1 y 9 1

i' - ;;j1

Y / I J K / L Fig. 32. .t1onnneboides latiguhernacula sp. n. A. Pharyngeal region. B. Anterior region showing stoma. C. Anterior region showing denticulate ridge. 1). Anterior region showing amphids dorsoventrally, E. Cuticle of anterior region. F. Anterior region showing Cheilorhabdions. G. Cuticle showing longitudinal ridges. H. Vulval opening (dorso%entral). I. Vulval region showing a pair ofdumb-bell shaped pouches, J. Female posterior region, arrow showing phasmids. K. L. Male posterior region showing spicules & gubernaculum, (Scale bars = 20µm).

ti Moaonchoides striatus (Butsehli, 1876) Goodey, I963 (Fig. 33, 34)

Measurements: In Table 17.

Descriptions

Females, Body almost straight upon fixation. Cuticle with transverse striations and

prominent longitudinal ridges. Amphidial apertures circular, at level of cheilostom.

Lip region continuous with body contour. Lips six, fused, each with a papilla. Stoma

longer than wide. Cheilostom wide. cuticularized. Cheilorhabdions arched inwards,

flaps bifid apically, extending beyond labial contour. Gymnoslorn anisofopfic, subventral walls longer than dorsal. S1egostom anisolopic and anisomorphic. Dorsal wall with a claw-like tooth, right subventral with a plate-like pyramidal tooth and left subventral with a denticulate ridge. Posterior part of stegostom tubular, longer than wide. Procorpus muscular, gradually expanding to a spheroid median bulb. Isthmus short. Anterior pharynx muscular, 64 — 75 % of pharyngeal length. Nerve ring encircling isthmus in the anterior half. Hemizonid slightly posterior to nerve ring.

Excretory pore further posterior to hemizonid, 75 — 84 °/ of the total pharyngeal length. Cardia small, distinct. Intestine with wide lumen.

Reproductive system amphidelphic, anterior branch on right and posterior branch on left side of intestine. Ovary long, oocytes arranged in several rows in germinal zone. Oviduct short, narrow, gradually expanding to form a spermatheca.

Spermatheca elongate, separated from the uterus by a constriction, containing sperms.

Uterus divisible into a long glandular and a short muscular part. Vagina muscular.

Vulval opening circular, pore-like. A pair of dumb-bell shaped pouches present at level of uterus. Rectum 1.0 — 1.5 anal body diam. long. Phasmids 1.1 — 1.6 anal body diam. posterior to anus. Tail long filiform, 1.3 — 2.9 times vulva-anus distance.

151 Males: Body smaller than females. Anterior part similar to that of females except for

four additional cephalic setae. Testis single, reflexed. Spicules free, ventrally arcuate,

1.7 — 2.1 anal body diams long. Gubernaculum simple, 47 — 56 % of spicules length,

provided with an appendage proximally and a sleeve distally. Genital papillae nine

pairs; three pairs precloacal, six pairs postcloacal. Genital papillae formula: v I, v2,

v3d / v4, ad, phasmids, (v5, v6, v7). pd. GP vl and v2 within the range of spicules.

Phasmids posterior to ad. GP v7 longer than v5 and v6. Tail divisible into a short

conoid part and a long filamentous part.

habitat and locality: Farmyard manure collected from the village, Chhedhar, Aligarh,

U.P., India.

Voucher specimens

Ten females and ten males on slides Mononchoides striauus /1-5 deposited in the nematode collection of Department of Zoology, Aligarh Muslim University,

Aligarh.

Rentarkc

The description and morphometric measurements of our specimens concur well with that of original specimens given by (l3utschli. 1876) Goodey, 1963.

Hom-ever, a slight difference was found in length of tubular part of stegostom (longer than wide vs as long as wide).

152 Table 17: Measurements (in µm) of Mononc/widesstrartus. Mean and S.D. given in parenthesis.

Characters Females (n = 10) Males (n = 10) L 905-1222(1052±94.7) 676-986 (833.3 t 91.5)

a 24.6- 37.4 (29.7f 3.2) 26.0-33.8(29.6* 2.4)

b 5.5-6.5(6.010.3) 5.33-6.96(5.97=0.5) c 2.2-3.1(2.8±0.2) 2.4 -3.1(2.8±0.2)

c' 13.2-20.2(17.1±2.2) 12.0- 18.1 (14.1 t 1.7)

V 38.5-42.1(40.4± 1.1) T - - 27.7-35.1(31.9+2.1) Maximum body width 30- 40(35.7 t 3.2) 24-35(28.2 t 3.1)

Lip width 13-19(15.8+1.9) 12-15(13.6±0.8)

Length of stoma 20-26(23.6+2.1) 17-20 (19.1 f 1.0)

Pharynx 57-195(174.2± 11.98) 121 - 151 (139.4+ 10.1) Excretory pore from anterior end 122-162 (139.5 t 11.3) 102-130(1157± 9.6) Hemizonid From anterior end 116- 153(135.1 f 12.2) 97-123(109.4 t 9.3) Nerve ring from anterior end 113-145(129.8 f 11.8) 85-110(100.2±7.4) Pharynx base to gonad 99-I80 (131.2 t 23.4) 85-165(129.0 f 24.2) Anterior gonad 100- 145 (1202 t 14.4) Posterior gonad 90-147(110.5±16.1) VBD 26-42 (55.4±4.7) Vulva 380- 515 (425.6 ± 40.3) Vulva- anus distance 145-312(246.8±41.3) Rectum 25-31(26.8+ 1.9) 23-31(27.5±2.5) Tail 295-425 (380.0 f 39.8) 239-380(299.9 t 45.1) ABD 21-25(22.3- 1.5) 19-25 (21.2 11.6) Phasmids from anus/cloaca 24 -37 (30.4 f 4.5) 25 -30 (28.4 + 1.7) Testis 215-310 (265.0 f 28.2) Spicules along axis 37 -43 (39.9 ± 2.0) Spicules along cord 30 - 36(33.3 ± 2.1) Gubernaculunt 18-24(20.3 = 1.5)

153 ir

i;

F

E, (

Fig. 33. 11ononrchoides .straitus. A. Entire female, B. Entire male, C. Anterior region showing amphids dorsoventralls. 1). Pharvngeal region, E. Cuticle showing longitudinal ridees. F. Female reproductive tract (Anterior), G. Male posterior region.

154

A , B C D ,,~,

} ~ f Litt 44

t ' 2

f 1 . 4 j L

i

f J ~

. t H K LM

i .. 4. .tlonortchoides straitus. A. Anterior region shop ing stoma. B. Anterior region showing amphids dorsoventrally, C. Anterior region showing Cheilorhabdions, D. Anterior region showing denticulate ridge, F. Anterior pharynx. F. Median bulb & posterior pharynx, G. Cuticle showing longitudinal ridges. II. Vulva) region, I. Female posterior region, J. Female posterior region showing phasmids. K. I.. Male posterior region showing spicufes & gubcrnacuIum, M. Male posterior region showing genital papillae & phasurids, (Scale bars = 20µm). Genus Oigolaineila Paramouov, 1952

Diagnosis: Body 0.7 — 1.5 mm long. Cuticle transversely and longitudinally striated.

Labial papillae long, bristle-shaped. Cheilostom with corona. Gymnostom forming a

very short, wide ring. Stegostom with claw-like dorsal and right subventral tooth.

Female reproductive system amphidelphic. Spicules short, free. Gubernaculum

plump. Tail long, filiform.

Type species: Diplogacter winchesi Goodey, 1929 = Oigolaimella lorigicauda (Claus,

1862) Fhrst von Lieven, 2003 — syn. Diplogaster longicauda Claus, 1862.

Oigolaimella brevigubernaculo sp. u, (Fig. 35, 36)

Measurements: In Table I8.

Descriptions

Females: Body almost straight on fixation, tapering towards both extremities. Cuticle

thin with weak transverse and longitudinal striations. Punetations in transverse row, discontinuous in lateral fields. Lip region flat. Lips six, each with a prominent labial papilla..Amphidial openings circular, behind the posterior end of stoma. Stoma slightly longer than than wide. Cheilostom cuticularized with cheilorhabdial flaps pointed anteriorly, extending beyond labial contour. Gymnostom much shorter than wide, anisotopic. Stegostomal walls anisomorphic, dorsal wall with a medium-sized claw-like tooth, right subventral with a small pointed tooth; left subventral with a ridge. Procorpus muscular, median bulb oval. Isthmus gradually expanding into a glandular basal bulb. Nerve ring encircling isthmus in anterior half. Hemizonid posterior to nerve ring, 75 - 78 % of pharyngeal length. Excretory pore posterior to hemizonid. 76-81 % of pharyngeal length. Cardia distinct. Intestinal lumen wide.

156 Reproductive system aniphidelphic, anterior on right and posterior on left side

of the intestine. Ovaries reversed, oocyces arranged in one or two rows- Oviduct short

with narrow lumen, expanded distally into a spermatheca. Spermatheca set off from

the uterus by a constriction, containing large-sized sperms. Uterus with clearly

divisible into muscular and glandular parts. Vagina muscular, one-third to one fourth

vulva) body diam. long. Vulva) opening oval. Rectum 1.2 — 1.5 anal body diam. long.

Phasmids 1.5 — 2.3 anal body diam. posterior to anus. Tail long filiform, whip-like, 19

— 27 anal body diam. long.

Males: Anterior end similar to that of females except for four cephalic setac. Testis

reflexed. Spicules small, 1.5 — 1.7 times cloaca] body diameter long, ventrally arcuate.

Gubernaculum small, keel-shaped, with a small pointed extension distally. Genital

papillae nine pairs; three pairs precloacal and six postcloacal. Genital papillae

formula: vl, v3d, v21 v4, ad, (v5, v6, v7), phasmids, pd. Genital papilla vi far anterior

to spicules when retracted; GP v2 located on the upper cloacal lip; pd considerably

posterior to v7. Phasmids at level of v7. Tail long filiform.

Type llabimr and locality: Farmyard manure collected from Aligarh, 11.P., India.

Type specimens

Holorype: Female on slides Oigolaimella brevigubernacula/6 deposited in the

nematode collection of Department of Zoology, Aligarh Muslim University, Aligarh.

Paratypes: Eight females and six males on slides (hgolaimella brevigubernacula /1-5

deposited in the nematode collection of Department of Zoology, Aligarh Muslim

University, Aligarh.

Diagonosis and relationship

The new species (7igolairnella brevigubernacula sp. it is characterised by a medium-sized right claw-like dorsal tooth, a small-sized pointed right suhventral tooth and a left subventral ridge, conspicuously large spermatozoa in the female

157 reproductive tract, keel-shaped gubernaculum with a small pointed extension distally

and nine pairs genital papillae.

The new species resembles 0. longicauda (Claus, 1862) apud Furst von

Lieven (2003) in having same body morphometrics, same body length, same length of

stoma. pharyngeal length, tail length, in having same shape and length of the spicules

and in number and arrangement of the genital papillae. However, the new species can

be differentiated from 0. longicauda in having more slender body (Max. body diam.=

16-20 pm vs 23 vs 32 pm), anteriorly located vulva (V= 36.3 —39.6% vs 55-62

%), position of phasmids in males from cloacal opening (21 — 25 pm vs 36-39 pm) and in the shape of gubernaculum (keel-shaped vs stirrup shaped).

Etymolo,D': The species is named after the small gubernaculum.

158 Table 18: Measurements (in µm) of Oigolaimella brevigubernacula sp. n. Mean and S.D. given in parenthesis.

Characters Holotype Females (n = 8) Males (n = 6) L 791 j 666 - 935 (839 t 90) 633-843(735±71) a 41.63 41.3-51.9(46.0±3.9) 45.9-56.2 (50.3±3.3)

b 7.0 6.5-8.1(7.3±0.5) 5.9-8.3(6.8±0.9)

c 2.68 2.6-3.2(2.8±0.2) 2.6-3.0(2.7±0.2)

c' 22.69 19.1-26.9(23.8±2.2) 17.7-30.0(21.2±4.3)

V 38.31 36.3-39.6(38.0±1.2) T 31.6-42.9(37.2±3.5)

Maximum body width 19 l6-20(18.3±1.5) 12-17(14.7±1.6) Lip width 8 7 - 9 (7.9 ± 0.6) 6-7(6.5±0.5) Length of stoma 8 7-9(8.1 ± 0.6) 6-8(6.7±0.8)

Pharynx 113 102-134(115±10) 93-117(108±8)

Excretory pore from anterior end 91 79 - 102 (91.2 ± 7.3) 75 - 92 (83.8 ± 5.5)

Hemizonid from anterior end 87 77 - 100 (89.0 ± 6.9) 72 - 89 (80.8 ± 5.3) Nerve ring from anterior end 79 68 - 88 (77.9 ± 6.4) 65 -81 (72.2 ± 4.9) Medianbulb(length) 15 14-17(15.3±0.9) 12-15(13.2±1.1) Median bulb (width) 12 11 - 13 (12.3 ± 0.7) 9- 10 (9.3 ± 0.5) Pharynx base to gonad 80 48 - 116 (90.3 ± 19.9) 56-147(83.8±30.3) Anterior gonad 110 90 - 125 (113.3 ± 10.7) Posterior gonad 105 82-130(109.1 ± 15.7) VBD 19 17-20(18.9± 1.2)

Vulva 303 261-359(319±34) Vulva - anus distance 193 185-250 (216 t 23.6)

Rectum 16 15-20(18.1 f 1.7) 15-16(15.7±0.5) Tail 295 210-350(305±41) 230-330(270±33.2)

ABD 13 11-14(12.8±0.8) 11-16(13.0±1.9) Phasmids from anus/cloaca 20 20 - 30 (24.9 ± 2.6) 21 - 25 (23.8 ± 1.5) Testis 230 - 335 (273 ± 36.6) Spicules along axis 16 - 25 (20.3 ± 3.5) Spicules along cord 13 - 1.5 (14.2 ± 0.7) Gubemaculurn 7- 9 (7.8 ± 0.7)

159 F

E

N y

J 1 .

Fig. 35. Oigalaiiiaella brei•igoberHuc•ula sp. n. A. Entire teinale, B. Entire male, C. Anterior region. D. Pharyngeal region, IV.. Female reproductive tract (Anterior). F. Female posterior region. G. Male posterior region.

160 BC ❑~ E

0

F

I°

. A

r

r( S 1

H 1

I- i_.. 36. OiL,,Iai,nc•llu hrL r uhc rvra~ u/a sp. n. A. Anterior phar\ nx. B. Anterior region showing stoma. C. D. Anterior region showing anteriorl pointed cheilorhabdial flaps. E. Anterior region sho%%insg amphids (lateral), F. Female reproducti\e tract (anterior branch), arrow showing sperms, G. Reflexed testis, H. I. Male posterior region showing spicules & gubernaculum, (Scale bars 20µm). Oigolal vela longicauda (Claus, 1862) Furst von Lieven, 2003 (Fig. 37. 38)

Measurements: In Table 19.

Descriptions

Females: Body almost straight on fixation, tapering towards both extremities. Cuticle

thin with weak transverse and longitudinal striations. Punctations in transverse row,

discontinuous in lateral fields. Longitudinal lines faint. Lip region flat, labial papillae

long setose. Amphidial openings circular, behind the posterior end of stoma. Stoma

wide. Cheilostom cuticularized with cheilorhabdial flaps pointed anteriorly, extending

beyond labial contour. Gymnostom much shorter than wide. Stegostornal alls

anisomorphic, dorsal with a medium-sized tooth, right subventral with a fine tooth;

left subventral with a ridge. Procorpus muscular, median bulb globular. Isthmus

gradually expanding into an elongate basal bulb. Nerve ring encircling isthmus in

anterior half, 64-71 % of total phanvngeal length. I lemizonid posterior to nerve ring,

68 — 81 % of total pharyngeal length. Excretory pore posterior to hemizonid, 71 — 74

% of the total pharyngeal length. Cardia distinct. Intestinal lumen wide.

Reproductive system amphidelphic. anterior on right and posterior on left side

of the intestine. Ovaries reversed, ooc\tes arranged in one or two rows. Oviduct short,

narrow, expanded distally into a spermatheca. Spermatheca set off from the uterus by a constriction, containing sperms. Uterus with clearly divisible into a short muscular and a long glandular part. Vagina muscular, more than one-third of the body diam. long. Vulval opening oval. Rectum 1.1 — 1.7 anal body diam. long. Phasrnids 1.7— 1.9 anal body diam. posterior to anus. Tail long filiform, 1.0 — 1.6 times vulva-anus distance.

.tfales: 13odh slender. Anterior end similar to that of females except for four cephalic setae. Testis reflexed ventrally. Spicules small, ventrally arcuate. Gubernaculum

162 large, stirrup shaped, without a distal sleeve. Genital papillae nine pairs; three pairs

precloacal and six postcloacal. Genital papillae formula: vi, v3d, v2/ v4, ad, (v5, v6,

v7), phasmids, pd. Genital papilla vi far anterior to spicules when retracted; pd

considerably posterior to v7. Phasmids at level of v7. Tail long filiform.

Habitat and locality: Farmyard manure collected from the District Baramulla,

Kashmir. Jammu and Kashmir.

Voucher specimens

Eleven females and nine males on slides Oigolaimella longicauda/l -6

deposited in the nematode collection of Department of Zoology, Aligarh Muslim

University, Aligarh.

Remarks

The description and measurements of our specimens agree well with 0. longicauda (Claus, 1862) aped Furst von Lieven (2003) in the morphometric values, in the general shape of body, in number and arrangement of genital papillae and in shape of spicules and gubernaculum. However, slight differences were found in the value of c' in females (20.7-26.3 vs 15.2 — 19.3) and position of phasmids from anus

(26 — 27 pm vs 29 — 48 pm in females and 23 — 25 pm vs 36 — 39 µm in males) and in length of gubernaculum (10— II pm vs 8-9 µm).

163 Table 19: Measurements (in µm) of Oigolaimella longicauda. Mean and S.D. given in parenthesis.

Characters Females (n = 11) Males (n = 9) L 893-1018(957±42) 760-941 (826±55) a 37.6 - 52.1 (44.4 ± 4.4) 40 - 47.7 (43.6 f 2.7) b 6.1-8.1(7.1±0.6) 6.3-7.5(6.8±0.4) c 2.5-3.3(2.9±0.2) 2.8-3.5 (3.1 ± 0.3) c' 20.7-26.3(23.3±1.7) 16-21.3(17.9±1.6) V 35.1 -40.9(38.9± 1.7) T 33-48.1 (43.4 ± 4.3) Maximum body width 18 - 25 (21.7 ± 2.1) 17 - 21 (19 f 1.2) Lip width I 10- 11 (10.4±0.5) 9- 10(9.3±0.5) Length of stoma 8 - 10 (9.5 ± 0.7) 7 - 10 (8.8 f 0.8) Pharynx 124-156(136±8.9) 113-132(122±6.2) Excretory pore from anterior end 93 - 103 (97.3 f 4.2) 82 - 96 (88.4 ± 4.4) Hemizonid from anterior end 91 - 11 1 (100.4 f 5.7) 79 - 93 (86.6 ± 4.9) Nerve ring from anterior end 84 - 100 (91.4 t 4.9) 74-84 (78.4 ± 3.3) Median bulb (length) 16-20(17.9±1.2) 14-17(15.2±1.1) Median bulb (width) 12 - 15 (13.4 ± 0.8) 10- 13 (10.9 ± 1.1) Pharynx base to gonad 82-140(115.4 ± 16.0) 40-135(76.3 ± 27.7) Anterior gonad 101 - 150 (122.6 f 17.4) Posterior gonad 107-. 155 (125.4 f 16.4) VBD 20-24 (22.2± 1.5) Vulva 333 -413 (373.2 ± 26.2) Vulva - anus distance 234 - 290 (261.3 ± 19.7) Rectum 17-24(20.8±1.95) 17-20(19.3±0.9) Tail 1 290-380 (323.2 f 27.5) 240-340(268± 29) ABD 12-16(13.9±1.2) 14-16(15±0.5) Phasmids from anus/cloaca 26 - 27 (26.5 ± 0.5) 23 - 25 (24 ± 0.8) Testis 265 - 435 (359 ± 50) Spicules along axis 20 - 24 (21.5 ± 1.5) Spicules along cord 15 - 20 (17.8 ± 1.7) Gubernaculum 10 - 11 (10.1 ± 0.3)

164 E

F G

Fig. 37. Oigoluimella lows iccuida. A. Entire female. B. Entire male, C. Anterior region ing amphids dorsoventrall~, P. Pharyngeal region. F. Female reproductive tract (Anterior). F. Female posterior region, G. Male posterior region.

165

"4

C

5',

4 N 44;

E

Fig. 38. Qigolainrella longicuuda. A. Anterior pharynx, B. Anterior region showing stoma & cheilorhabdial flaps. C. Anterior region showing arnphids (dorsoventral)" 1). Female posterior region, F. Male posterior region sho%%ink: spicules & gubernaculum. (Scale bars 20pm). Genus Pristionchus Kreis, 1932

Diagnosis: Body length ranged 0.5- 2.5 nom. Cuticular annulations generally resolved

in fine punctuations. Amphids small. Buccal cavity consisting of one chamber and

shows dimorphism in each sex (steno- and eurystomous morphs). Cheilostom in

eurystomous morph with six plates. Metastom in stenostornous form with a triangular dorsal tooth and a blunt right-subventral tooth, in eurystomous form with a claw-like dorsal tooth and a similar right-subventral tooth. On left-subventral sector of both

morphs, a row of denticles present. Female gonads amphidelphic. A narrow bursa often present. Gubernaculum with a paired and an unpaired ventral process. Tail conoid, finely attenuated.

Type species: Pristionchus longicaulotus Kreis, 1932 = P. lheritieri (Maupas, 1919)

Paramonov, 1952 — syn. Diplogaster lheritieri Maupas, 1919.

Pristionchus slercoraris sp.n. (Fig. 39, 40)

Measurements: In 'Table 20.

Descriptions

Females: Body straight or slightly arcuate upon fixation. Cuticle with faint annulations. Lip region round continuous. Lips six. each bearing a small outer labial papilla. Amphidial opening oval at the level of anterior end of the dorsal tooth. Stoma spacious, both steno- and eurystomatous types, with stoma length 8 — 9 µm.

Cheilostom represented by a thin-walled arch. Gymnostom anisotopic with dorsal wall shorter than subventrals. Stegostom anisitopic and anisornorphic, dorsal wall with a large class-like tooth, right subventral \%all with a retrose triangular tooth and left subventrats with a serrated ridge. Anterior pharynx muscular, 57 — 65 % of the pharyngeal length. Median bulb muscular with oval-shaped strong valve-plates.

167 Isthmus long expanding posteriorly to a globose, glandular basal bulb. Nerve ring

located at middle of the isthmus. 65 — 79 °%o of pharyngeal length. Excretory pore near

basal bulb. Hemizonid just anterior to the excretory pore, 79 — 83 % of total pharynx

length. Cardia large elongate. Intestinal lumen wide.

Female reproductive system amphidelphic. Anterior genital branch positioned

to the right of intestine and posterior to the left. Ovaries reflexed, flexures extending

beyond vulva. Oviduct long, narrow. Spermatheca oval, not set-off from uterus,

containing sperms usually. Uterus divisible into a long glandular and short muscular

parts. Vulva transverse, occasionally protruding. Rectum length approximately equal

to 1.1 — 1.4 times anal body diameter. Phasmids 1.2 — 1.7 anal body diam. posterior to

anus. Tail acute and long, tapering gradually from anus to the terminus.

Males: Body J-shaped when heat-relaxed. Stoma as in females. Anterior end similar

to that of the females except for four additional cephalic setae. Testis single and

reflexed ventrally. Spicules slender, arcuate. cephalated, I.5 — I.6 anal body diam.

long, with distal tip slightly curved anteriorly. Gubernaculum boat-shaped with a

medial dorsal convex. proximally with an anteriorly directed needle-like extension

and distally with a sleeve, 48 — 50 % of the total spicules length. Genital papillae

eight pairs: two pairs preclaocal and six pairs postcloacal. Genital papillae formula:

v 1, v3d/v4, ad, phasmids, (v5, v6, v7), pd. GP v 1 beyond the range of spicules. GP v2

absent. GP v4 just below the lower cloacal lip. Phasmids posterior to ad. GP v7 and

pd almost at the same level

Type habitat and locality: Farmyard manure collected from the District Baramulla.

Kashmir. J & K.

TT pe specimens

I

168 Holotype: Female on slides Pristionchus stercoraris sp. n./5 deposited in the

nematode collection of Department of Zoology, Aligarh Muslim University, Aligarh,

Paratypes: Seven females and three males on slides Pristionchus stercoraris sp. n./l-

4 deposited in the nematode collection of Department of Zoology, Aligarh Muslim

University. Aligarh.

Diagonosis and relationship

The new species Pristionchus stercoraris sp. n. is characterized by faint

annulations on cuticle, steno- as well as eurystomatous types of stoma, thin-walled

cheilostom. anisitopic and anisomorphic stegostorn, a large claw-like dorsal tooth, a

retrose triangular right subventral tooth, a serrated ridge on left subventral wall, long

isthmus, long elongate cardia, amphidelphic female reproductive system, slender

spicules with anteriorly curved distal tip, a boat-shaped gubernaculum with a medial

dorsal convex: proximally with an anteriorly directed needle-like extension and

distally with a sleeve, and eight pairs of genital papillae.

The new species ressemble Pristionchus lheritieri (Maupas, 1919) Paramonov,

1952 in general body morphometrics, same body length, and amphidelphic female

reproductive system. However, the new species differs from P. lheritier in greater a

value (28.9-33.2 vs 16 — 17 in females and 25.6-30.3 vs 13-20 in males), greater

b value in males (6.4 — 6.8 vs 4.7 — 6.3), slightly smaller c value (3.2 — 3.7 vs 3.8 — 5.3

in females and 4.2-4.4 vs 5 — 9 in males), greater c'value (13.8— 19. 0 vs 6— 10 in

females and 7.9 -- 8.4 vs 3 —4 in males), in position of vulva (39.7-41 % vs 44 — 50

%). smaller spicules and guhernaculum (29 — 30 pm vs 35 — 50 µm and 14 — 15 pm vs

16 — 22 µin, respectively ). shape of gubernaculum (boat-shaped. a sharp anteriorly directed needle-like extension proximally vs proximalk rounded) and in number of

169 genital papillae in males (eight pairs: two preclaocal, six postcloacal vs nine pairs: three preclaocal, six postcloacal).

Etymology: The name of the species is derived from the Latin word of farmyard manure from which it was collected.

170 Table 20: Measurements (in µm) of Pristionchus stercoraris sp. n. Mean and S.D. givcn in parenthesis

Characters Holotype Females (n°7) Males (n=3) L 953 804-1042(958+71) 626-728(681 f42) a 32.86 28.9- 33.2 (31.3 f 1.5) 25.6-30.3(27.7 f 1.9)

b 7.75 7.0-9.3 (7.9 t 0.7) 6.4-6.8(6.6+0.2) c 3.29 3.2-33(3.4 t 0.2) 4.2-4.4 (4.3 t 0.1)

17.06 13.8-19.0(16.3= 1.6) 7.9-8.4(8.2±0.2)

V 39.66 39.7-419(40.6t0.9) T 47.9-57.3 (53.1 ± 3.9) Maximum body width 29 26-35(30.7 t 2.7) 23-27(24.7f 1.7)

Lipwidth 13 12-13(12.6±0.5) 10-11(10.3±0.5)

Length of stoma 8 8-9(8.3±0.5) 6-7(6.7 t 0.5)

Pharynx 123 107-130(121.1±7.5) 98-107 (103.3 f 3.9) Nerve ring from anterior end 81 77-86(82.6+3.1) 73-75(74.0±0.8)

Median bulb (length) 19 15-19(17.4+1.4) 14-15(14.7±0.5)

Median bulb (width) 16 12-17(15.3 t 1.9) 12-13(12.3±0.5) Pharynx base to gonad 110 75-129(100.1=16.1) 30-78(57.0120.1)

Anterior gonad 145 115-235(168.1 f 35.4) Posterior gonad 165 125-210(169±23.6)

VBD 29 26-37(31.1±3.1) Vulva 378 319-437(389.4+34.7) Vulva - anus distance 285 235-325(286.9 t 25.8) Rectum 20 17-23(20.3+1.8) 20-22(20.7+0.9)

Tail 290 250-315(282.1+ 21.5)' 150-165(158.3+6.2)

AI3D 17 15-20(17.411.8) 19-20(19.3+0.5)

Phasmids from anus/cloaca 25 23-25(24.1 f 0.8) Testis 300-395 (362+44)

Spicules along axis 29-30(29.7+0.5) Spicules along cord 25 - 26 (25.3 ± 05) bernaculumGu 14-15(14.7±0.5)

171 [;1

IN I

-'H

., 13 t5fl 1 iiii

C-G------20 gm

Fig. 39. Pristiww/:us .siercorurzs sp. n. \. Entire female. B. Entire male, C. Pharyngeal re,-non, D. Female reproductive tract (Anterior). F. Female posterior re2ion. F. Male posterior region. G. Spicules & Gubernaculum.

172

A C D E

I

B 4

K

I ,

.74-1~~A v

7 rj

-gap "A

OL

L t H j

Fig. 40. Pristhinehu.v sI0'Lr1Fi, sp. n. A. 13. Anterior region showing stoma. C. D. Anterior region showing cheilorhabdions. F. Anterior region showing amphids (dorsoventral). F. Anterior pharynx. G. Median bulb, isthmus, basal bulb & cardia. II. Ovaries showing overlap. I. Vulval region. J. Female posterior region. K. Female posterior region showing phasmids

(dorsoventral). L. Male posterior region showing spicules & gubernaculum, (Scale bars = 20jim). Family Tylopharyngidae Filipjev, 1934

Diagnosis: Amphids behind lips, transverse. Mouth cavity very long and thin, similar

to the Tylenchida. Cheilo- and gymnostom very short, tube-like, with two basal

(mobs. 'teeth small. Pharynx with muscular anterior and non-muscular posterior

section. Female gonads paired. Bursa absent. Tail long in both sexes.

Type subfamily: Tylopharynginae Filipjev, 1934

Genus Tvlopharynx de Klan, 1876

Diagnosis: Body 0.6 to 1.6 mm long. Cuticular staitions divided by longitudinal

striae. Labial papillae minute. Mouth cavity resembling a Tytenchus-like stylet.

Cheilo- and gymnostom very small, the former not divided into plates. Metastom

bearing a very small dorsal and a similar right subventral tooth; left subventral section

unarmed. Telostom very narrow, six to ten times as long as wide, with two

subdorsally originated basal knobs. Pharynx more slender than in other diplogastrids,.

median bulb strong, spherical. Female amphidelphic. Bursa absent. Nine pairs of

genital papillae. Spicules arcuate. Gubernaculum small. Tail of each sex long,

fl! iform.

Type species: Tylophorynr foetida (BUtschli, 1874) Goodey, 1928 — syn. Aphelenehus foeridus Butschf, 1874

Tylopharynxfoetrda (Butschli, 18?4) Goffart, 1930 (Fig. 41, 42)

Measurements: In Table-21.

Descriptions

Females: Body almost straight after fixation. Cuticle faintly annulated and bearing shallow longitudinal striae. Lip region appears smooth, continuous with the body

174 contour or separated from the rest of the body by a slight depression. Cephalic

framework moderately developed. Lips Six, amalgamated, each bearing a minute

labial papilla. Amphidial fovea rarely distinct in lateral view, goblet-shaped.

Amphidial apertures slit-like, just below the anterior opening, more or less obscure in

lateral view. Stoma 1I — 13 µtn long. Chielostom and gymnostom wery short.

Stegostom long, tube-like and narrow anteriorly with a dorsal tooth and a right

subventral tooth and posteriorly with two subdorsal knobs at its base. Knobs 1.5 — 2.0

gm in diam. Anterior pharynx cylindrical, muscular, 55 — 58 % of total pharynx

length. Median bulb measuring 12 — 15 µm long and 9 — 11 pm with crescentic

longitudinal valves. Isthmus narrow, expanding into a glandular basal bulb. Nerve

ring surrounding isthmus at 71 — 106 µm from anterior end. Excretory pore located

posterior to the hemizonid, opposite to the basal bulb. Hemizonid just anterior to

excretory pore, 75 — 83 % of the total pharyngeal length. Cardia small but distinct.

Intestine with wide lumen.

Female - reproductive system amphidelphic, with antidromously reflexed

ovaries. Anterior gonad on the right side and posterior gonad on the left side of

intestine in all females. Oocytes arranged in two or more rows. Oviduct short and

narrow. Spermatheca slightly demarcated from uterus. Uterus divisible into a large

glandular and a narrow muscular part. Vulva small pore-like. Vulva-anus distance 8 —

10 vulval body diams long. Rectum 1.0 — 1.3 anal body diam. long. Tail conical.

tapering smoothly to a long, filiform terminus, 1.2 — 1.5 times vulva-anus distance

long. Phasmids conspicuous, 1.4 — 1.7 times anal body diam. posterior to anus.

Ala/es: Similar to females in most respects. Body straight after fixation except for the ventrall\ curved cloaca! region. Lip region with four cephalic sensilla. in addition to the six labial sensilla also found in the females. Testis single, ventrally reflexed,

175 located on right side of intestine. Spicules ventrally arcuate. 1.9 — 2.0 times anal body

diam. long, proximal end cephalated and distal end with a pointed digitate terminus.

Gubernaculum simple, thin, 35.7 — 37.9 % of total spicule length. Nine pairs of

genital papillae: three pairs precloacal, six pairs postcloacal. Genital papillae formula:

(v1, v2), v3d / v4, ad, ph, (v5, v6, v7), pd. GP v2 lies adjacent to GP v1. GP v3d just

above or at the level of upper cloacal lip. GP vl, v2 and v3d in the spicular range. GP

v4 slightly posterior to the cloaca. GP v5,6,7 are grouped close together posterior to

phasmids. Phasmids located 1.2 to 1.3 anal body diam. from anus. Bursa absent.

Rectum 1.2 — 1.3 anal body diam. long. Tail in two parts: a short conoid part and a

long filamentous part.

Habitat and locality: Farmyard manure collected from district Baramulla, Kashmir

J&K. India.

Voucher .specimens

Six females and three males on slide TmlopharvHvfoetida/l-4: deposited in the

nematode collection of Department of Zoology. Aligarh Muslim University, Aligarh.

Remarks

There are only two species representing the genus TpIopham•m, namely T.

foetida (Butschli, 1874) Goffart. 1930 and T. clariamphida Wu et al. (2001). T.

foetida has been reported from different parts of Europe and T. clariamphida was

collected from China. This is the first report of the genus Tylopharynx from India.

Measurements and descriptions of our specimens agree well with that of the original

specimens of T. foetida de scribed by Butschli, 1874. However, differences were

found in having shorter stoma (l 1 — 13 µm vs 14— 15 µm), longer tail (c = 2.7 — 3.09

vs 3.4 - 4.8). shorter anal body diam. (c' = 18.2 — 20.4 vs 9.8 — 15). slightly smaller

anterior pharynx (59 — 70 p.m vs 72 — 81 µm), and in position of excretory pore

' (opposite to basal bulb vs opposite to isthmus).

176

Table 21: Measurements (in µm) of Tylopharynxfoetida. Mean and S.D. given in . parenthesis.

568- 813 (726.3 t 57.2) 599-665(635_33

la 30.6-35.2(33.08x1.9) 33.8-36.9(35.3*

5.6-6.7 (6.2 t 0.4) 5.1 -5.2(5.15±0

c 2.7-3.09(2.9 t 0.1) 3.2-33(3.3+0.

C? 18.2-20.4(19.3+0.9) 12.9-13.7(13.31 V 38.6-42.1 (39.8+1.1)

37.9- 39.10 (38.7 t 0.5)

Maximum body 19- 25 (221 1.8) 17-19(18+0.8)

Lip width 5-6(5.7=0.5) 5-6(5.710.5)

11-13(12.2±0.7) II- 12 (11.7 t 0.5)

v-rg 179+u 117-130(123.3±5.3) ae- IuLpLx 1.3) 89-95(917174)

71-106(82± 11.7) 12-15(12.8±1.07) 13-15(14*0.8)

9- I1 (10.2±0.7) 10-11(10.3±0.5)

Pharynx base to gonad 53-71 (61.815.5) 69-75(71.3 12.6)

Anterior gonad 92- 133(110.3 f 16.1) 80-106 (90.5 f 9.8) 20-24(21.5± 1.4) 263 - 319 (289.3125.$ anus 63-216(186.2± 19.9E

13-16(14.7= 1.3) 17-20 (18.7 f 1.3) ?35 - 285 (250.8 + 16.4, 30-205 (195 + I0.!

12-14(13X0.8) I4-15(14.7+05) 19 -21 (20±0.71 17-20(18.3 f 1.3) 7-260(245.7±13.

28 -30 (29 ± 0.8)

22-25 (237 f 1.3)

10-11 (10.70.5)

177 17

oLIJ JO I

Ji

F

Fig. 41. Tr1ophuriivJoetida. A. Entire tmaIe. 13. Fntire male. C. Anterior end, D. Pharvneal region. F. Female reproductive tract (Anterior), F. Female posterior region. G. Male posterior region.

178 A C

I E G

i~ )

. .

H

Fig. 42. Tvlophart-nx f etida. A-D. Anterior region showing stoma, arrow showing subdorsal knobs. E. Anterior pharynx. F. Posterior pharynx. G. Female reproductive tract (anterior branch), H. Vulval region. I. Female posterior region, J. K.. Male posterior region showing, spicules & gubernaculum, (Scale bars 20 µm). SUPERFAMILY CYLINDROCORPOROIDEA Goodey, 1939

Diagnosis: Stoma cylindrical, very long and narrow, tubular approximately one-

fourth of total pharyngeal length, stoma without true teeth. Anterior pan of pharynx

well developed, muscular, cylindrical, procorpus as wide as median bulb. Bursa

present.

Type family: Cylindrocorporidae Goodey. 1939

Other family: Lougibuctidae Poinar, Jackson, Bell & Wahid, 2003

FAMILY CYLIN'DROCORPORIDAE Goodey, 1939

Diagnosis: Stoma very long, narrow tube-like, one-fourth of total pharyngeal length

or even longer. Pharynx divisible into two parts: anterior cylindrical and strongly muscular; posterior pan long and glandular. Amphids on lateral lips. Female reproductive system amphidelphic or monoprodelphic. Female tail long, whip-like.

Male with single testis, bursa present, often rudimentary: pclodcran or leptoderan, nine or ten pairs of genital papillae. Spicules paired, gubernaculum present.

Genera: Goodeyus Chitwood, 1933

Myctolarmus Cobb, 1920

. ,lyctolarn ellus Andrassy, 1984

Protocylindrocorpus Ruhm, 1959

Genus Mycaolaimu.s Cobb, 1920

Diagnosis: Head composed of six lips each hearing a papilla. Stoma long, narrow and tubular. Cheilorhabdions present. Pharynx consisting of a cylindrical precorpus ans corpus with rounded ends and tri-radiate lumen, followed by narrow isthmus and terminal bulb without valve plates. Female with pre-equatorial vulva, gonads paired, opposed and reflexed. Male with single testis and nine pairs of genital papillae. Bursa

180 absent, if present rudimentary. Spicules paired, tapering and blunt. Gubeniaculum

present.

Type species: Mycmlaimus pellacidus Cobb, 1920

Mrcto/aimus papillatus sp. n. (Fig. 43, 44)

Measurement: In Table 22.

Descriptions

Females: Body medium-sized. straight when fixed, tapering at extrernities, more

towards the posterior end. Cuticle thin with prominent longitudinal lines and fine

transverse striations. Lip region continuous with the body, 2 in high and 6-7 pm

wide. Lips six, protruding, each with a small papilla. Amphidial apertures

indiscernible. Stoma tubular, narrow and flexible. Cheilostom short, walls

cuticularized, wider than gymnostom. Gymnostom 5-6 times lip-widths long.

Slegostorn short, without tooth and denudes, 4-6 gm long. Pharynx consists of an

elongate cylindrical, muscular bulb with valve plates, 3.0-3.5 times longer than wide.

Isthmus long, slender, gradually expanding posteriorly to form a basal bulb. Basal

bulb pyriform and glandular. Corpus 58-61% of pharynx. Nerve ring encircling

isthmus in middle. Dierids at the level of basal bulb. Ilemizonid slightly posterior to

nerve ring. Excretory pore posterior to hemizonid. Cardia muscular and well

developed. Intestinal cells large, granulated, lumen wide.

Reproductive system amphidelphic, anterior branch on right and posterior on

left side of intestine. Ovaries long, reflexed, extending almost upto vulva . Oviduct

long, and narrow, expanding posteriorly to form a spemtatheca. Sperniatheca set-off

from uterus by a constriction usually containing spermatozoa. Uterus with long glandular and short muscular parts. Vagina muscular with two pairs of vaginal

181 pouches, one pair on either side of the vulva. Vulva a transverse slit, vulval lips

slightly raised. Phasmids 20 - 29 pin from anus. Tail elongate eonQid with a pointed

terminus.

Males: Body medium-sized, ventrally curved at posterior region when fixed. Anterior

region similar to that of females. Testes single, ventral in position, flexure on right

side. Spermatogonia arranged in multiple rows in the germinal zone. Spicules slender

1.3 - 1.6 anal body diam. long, slightly ventrally arcuate with capitate proximal and

blunt distal ends. Gubemaculum straight, plate-like; distal end finely pointed with sleeve. Genital papillae nine or ten pairs; three (or four) pairs precloacal, six pairs postcloacal. Formula for genital papillae: vi, v2, v3d / v4. ad, phasmids, (v5, v6, v7), pd. The additional (tenth) when present, anterior to vl. Bursa absent, but cuticle slightly inflated in the vicinity of (v5-7) group. Tail consists of a short conoid part and long filamentous part with pointed end.

Type habitat and locality: Collected from farmyard manure. Kanispora, l3aramulla.

Kashmir, India.

Type specimens

Holotype: Female on slides Alyctolainms papifalus sp. n./3, deposited in the nematode collection of Department of Zoology, Aligarh Muslim University, Aligarh,

U.P.

Paratypes: Ten females and ten males on slides Mycto1nimus papillatus sp.n./I-10 deposited in the nematode collection of Department of Zoology, Aligarh Muslim

University, Aligarh.

Diagnosis and relationship

Afyctolaimus pupiIlazus sp. n. is characterized by fine transverse and longitudinal striations, nine or ten pairs of genital papillae in males, slender and arcuate spicules, gu6emaeulmrn with distal sleeve, and a long filiform tail.

182 Myctolaiinus papillarus sp. n. closely resembles M. macrolaimus (Schneider,

1866) Andrassy, 1984 and M neolongistoma Hussain et al., (2004). The new species can be differentiated from both the related species in structure of gubernaculum ( with sleeve vs without sleeve), bursa (absent vs rudimentary) and in the arrangement of the ten genital papillae (4 pre- and 6 postcloacal vs 3 pre- and 7 postcloacal). It further differs from M. macrolaimus in the presence of vaginal pouches (absent in M macrolaimus). Further differences from Al. neolongistoma are in body length of males

(623-915 vs 433-461 gm), and in position of precloacal genital papillae (v2 and v3d in spicular range vs vI, v2 & v3d in spicular range).

Etti•molog': The name of the species is derived from the papillae.

183 Table 22: Measurements (in gm) of Myctolaimus papillatus sp. n. Mean and S.D. given in parenthesis.

Characters Holotype Females (n=10) Males (n=10) L 883 688-974(785±77) 623-915(732±50) a 31.5 26.2-31.5(28.8± 1.5) 24.6-29.5(27.1±1.4)

b 7.1 5.7-7.1 (6.2 ± 0.4) 4.7-7(5.2±0.6) c 5.4 4.7-6.1(5.4±0.45) 14.1-19.5(17.9±2.1) c' 9.2 7.2-10.6(9.0± 1.1) 1.6-2.1(1.9± 0.2)

V/T 47.1 45.9- 49.6(47.5 t 1.0) 53.8-71.3 (61.6 ± 5.3) Lip width 9 6 - 7 (6.2 ± 0.4) 5 - 7 (6 ± 0.4)

Length of stoma 45 37-49(41 ± 3.3) 37-45 (41 ± 2.7) Anterior Pharynx 83 81 - 97 (86 ± 4.3) 75 - 95 (84.6 ± 5.3)

Posterior Pharynx 42 55 - 70 (59 ± 3.9) 55-80 (61.6 ± 7.2) Length of Pharynx 125 138-167(146 ± 8) 130-175 (146 ± 12)

Excretory pore from anterior end 126 110-143(124 ± 9) 111- 126(120±4) Hemizonid from anterior end 118 105 - 139 (115 ± 9) 107 - 120 (1 14 ± 4.6) Nerve ring from anterior end 103 97- 127 (105 t 8) 99 - 107 (103 ± 3) Median bulb length 44 46 - 56 (50 ± 3) 43 - 53 (47.8 ± 2.6) Median bulb width 14 13-18(15± 1) 12-15(14± 1) Anterior gonad length 118 105-163(118 ± 13) Posterior gonad length 112 94 - 159 (110 ± 17.7) Vulval body diam. (VBD) 30 23 - 38 (27 ± 3.9) Vulva-anus distance 190 200 - 344 (240 ± 40) Rectum (length)/cloaca 21 14-23 (18 ± 3) 19-23 (20 ± 1) Tail length 165 150-200(178 ± 14.9) 110-145(126 ± 10) Anal body diam (ABD) 18 14-22 (17 ± 2) 17-24 (19 ± 2.5) Phasmids from anus/ cloaca 20 20-29 (23 ± 3) 16-22 (19 ± 1.8) Testis length 245 - 470 (346 ± 63) Spicules length along axis 23 - 31 (27 ± 2.2) Gubemaculum I I - 15 (13 ± 1.5)

184 D [S F

\\ \ \\(\ \\

1141

\! E

Fig. 43. Mj'ciolaunu.s papillulus sp. n. A. Entire female, B. Entire male, C. Pharyngeal region. 1). Female reproductive tract (Anterior), F. Female posterior region. I. G. Male posterior region.

185 It

H F

oj-

J K

thcI(,I(II,nus Imp/ \ Pharyngeal region: 11 Posterior genital hraiii. Ii sho"ing, sphincter: D & L. Vulval region showing vaginal pouches (Lateral & Ventral view): F. Vulval opening, (ventral): G. Female posterior region showing phasmid (dorsoventral): II. ton2itudinal lines: I. Male posterior region showing musculature; J. (iubcrnaculum: K. Spicules. (Scale bars 20im). Myciolaimus oreodorus sp.n. (Fig. 45, 46)

Measurements: In Table 23.

Descriptions

Females: Body medium-sized, straight or slightly ventrally curved when fixed.

Cuticle thin with prominent 51 longitudinal lines and Uric transverse striations. Lip

region continuous with the body, 2-3 pm high and 7-9 µm wide. Lips six protruding,

each with a single small papilla. Amphidial apertures indiscernible. Stoma tubular,

narrow, and flexible. Cheilostom short, cuticularized, wider than gymnostom.

Gymnostom tubular, 4-5 lip-widths long. S'tegostom short, without tooth or denticles, surrounded by 4-5 µm long pharyngeal tissue. Pharynx consists of an elongate cylindrical, muscular bulb with valve plates, 3.0-3.7 times longer than wide. Isthmus long, gradually expanding posteriorly to form a pyriform basal bulb. Corpus about 60-

68 percent of pharynx. Nerve ring encircling isthmus in posterior half or at 73-82% of pharyngeal length. Hemizonid posterior to nerve ring. Excretory pore posterior to hemizonid. Cardia well developed, 5-6µm long . Intestinal lumen wide with large granulated cells.

Reproductive system amphidelphic, anterior branch on right side and posterior branch on left side of intestine. Ovaries long, reflexed with well developed sphincter muscles, usually extending up to or beyond vulva. Oocytes arranged in multiple rows in germinal zone. Oviduct short and narrow, expanding posteriorly to form a spermatheca. Spermatheca either continuous or slightly set off from uterus by a constriction and usually containing a large oval spermatozoa. Uterus with long glandular and short muscular parts. Vagina muscular with two pairs of vaginal pouches, one pair on either side of vulva. Vulva a transverse slit, vulva] lips without protuberance except in few specimens. Rectum short, about one anal body diam. long.

187 Tail elongate conoid with a pointed tip. Phasmids about 0.7-1.2 anal body diam.

posterior to anus.

Mates: Body smaller than females. Anterior region similar to that of females. Testis

single, ventral in position, flexure on right side. Spermatogonia arranged in one Cr

more rows in the germinal zone. Spicules paired. slender, free, 1.2-1.9 anal hody

diam. long, slightly ventrally arcuate with capitate proximal and blunt distal ends.

Velum moderately developed. Gubernaculum simple, plate-like; proximal end blunt,

distal end finely pointed, with sleeve. Genital papillae nine pairs; three pairs

precloacal, six pairs posteloacal. A ventromedian papilla present on the anterior cloaca) lip. Formula for genital papillae: vi, v2, v3d / v4, ad, ph, pd (v5, v6. v7).

Posterior dorsal (pd) genital papilla located anterior to the group of v5-7. Bursa

Icptoderan, very narrow, almost sticking to the lateral body walls. Tail short conoid with a small tail spine.

Type habitat and locality: Dry inflorescence of bottle palm, Oreodoxa regia collected from the Department of Zoology, Aligarh Muslim University. Aligarh, India.

Other population collected from rotting wood, Keirak, Thoubal, Manipur, India. type specimens

Holotvpe: Female on slides Myelofaimus oreodoxus sp.n./l, deposited in the nematode collection of Department of Zoology. Aligarh Muslim University, Aligarh,

U.P.

Paratypes: Ten females and ten males on slides Myctolaimua oreoduxus sp.n./2-8 deposited in the nematode collection of Department of Zoology, Aligarh Muslim

University, Aligarh.

Diagnosis and relationships

.Myciolaimus oreodoxus sp. n. is characterized by a thin cuticle with longitudinal lines, ovaries with well developed sphincter muscles and long conoid

188 female tail. Males with nine pairs of genital papillae (3 pre-cloacal and 6 post- cloacal), slender and arcuate spicules, gubernaculum with distal sleeve, narrow leptoderan bursa and a short conoid tail with small spine.

The new species closely resembles M kishtwarensis Hussain el al. (2004), M. curzii (Goodey, 1935) Andrassy, 1984, M. analophorae Kanzaki & Futai (2004). M. oreodoxus sp. n. can be differentiated from all the related species by the structure of the ovaries (with sphincter vs without sphincter), and in structure of gubernaculum

(with sleeve vs without sleeve). M. anoplophorae and Al. kishtwarensis also differ in having smaller male tails (c = 14.1-19.5 vs 8.3-11.9; c =14.1-19.5vs 5.5-8.7 respectively). Additional differences from Al. curzii, M. kishtwarensis and M. rifjlei is in the structure of the cuticle (with longitudinal striations vs without longitudinal striations), and position of pd (anterior to v567 group vs posterior to v567 group in M. curzii & M. kishtwarensis).

Etymology: The name of the species is derived from the name of the host plant from which it has been collected.

189 Table 23: Measurements (µm) ofMyctolaimus oreodoxus sp. n. Mean and S.D. given in parenthesis.

Characters Hololype Females (n=10) Males (n=10)

L 837 755-973 (852364) 593-786(672±50)

a 32.2 262- 31.5 (28.8* 1.5) 24.4-28.5(263±1.2)

b 5.7 5.7-7.0(6.230.4) 4.6-6.9 (5.2 t 0.6)

c 4.2 4.7-6.1(55x0.5) 14.1-19.5(17.912.2) c 11.7 7.2-12(9.0 ± 1.0) 1.6-2.1(2 ± 0.2) VT 44.5 46-49.5(47.5 ± 1.0) 54-71 (61.5 t 5.5)

Lip width 7 7-9(8.5 ± 0.5) 7-9(7.5±0.5) Length of stoma 41 41-50(45±3) 40-48(44.5±2) Anterior pharynx 87 80-96(88 f 5.5) 72-85 (80 i 4) Posterior pharynx 60 42-60(50 3 5) 42-62(52.5 ± 5)

Leagth of pharynx 147 125-149037± 8) 103-147 (129 ± 11) Excretory pore from anterior end 121 125.5 ± 9.5(116 -137) 106- 127 (118 ±7) Hemizonid from anterior end 115 110- 131 (119 ± 7.5) 101- 120 (112 ± 5.5) Nerve ring from anterior end 102 94 - 161 (111 ± 18) 94-106(98 ± 5) Median bulb length 50 42-53(465± 3.5) 35-45(40 ± 3.0)

Median bulb width 14 12-15(13.53 1) 10-14(12.53 1) Anterior gonad length 120 101-167 (132 ± 22)

Posterior gonad length 122 80- 181 (129 ± 29.5)

Vulva) body diam. (VBD) 27 28-41(32 ± 3.7) Vulva-anus distance 265 137 -190(161 i 14) Rectum(length)/cloaca 19 20-25(21 f 2) Tail length 200 131-180(157 ± 15) 34-42 (38 i 2.7) Anal body diam. (ABD) 17 15-22(17.5 t 2.5) 17- 21 (19 ± 1.2) Testis length 330-545 (417 f 64) Spicules length 23 -28 (25.7 f 1.5) Gubernaculum I1 - 12 (12 ± 0.5)

190 C I1\

F 31,

D- -.20 urn 20tm (; A,B -4lOOJLtnl 20 .tln

Fig. 45. M c1o1unusii oreodoxus sp. n. A. Entire tiiia1c, B. Entire male, C. Pharyngeal region, U. Female reproductive tract (Anterior). E. Female posterior region. F. Male posterior region (dorsoventral), U. Male posterior region (lateral).

191 ED F

H J F

L M'

Fig. 46. ifvct,,i01?11 i. A. l'harngeal rLi.ioii. B. Mma C Part ol jJ ditmim, sphincter: D. Anterior gonad shoing sphincter: F. Vulval region shoing muscles: F. Vulval region showing vaginal pouches: G. Vulva (Ventral ie"%), H. Longitudinal lines: I. Female posterior region showing phasmid (dorsoventral): J. Male posterior region showing musculature: K. Spicules & guhernaculum,- L & M. Male posterior region showing tail and genital papillae. (Scale bars 20im).

Embryogenesis

The intra-uterine development was not found in Alyctolaimus oreodoras sp. n.

Not more than two eggs were present in the uterine tract of gravid females. Eggs laid

were usually single celled, sometimes eggs with two cells were also laid. The process

of egg-laying was found to occur within a minute.

Egg: Eggs were ovoid, elongated, smooth-shelled with dense granular cytoplasm and

a fairly large nucleus and measured about 53-67 pm in length and 23-28 µm in width.

A polar body is usually visible between the cytoplasm and the vitelline membrane.

Fertilization: Sperm penetration takes place as soon as the mature oocyte reaches the

uterus from the oviduct. The point of the sperm entry marks the posterior side of the egg. The sperm and egg pronuclei were situated at the opposite poles. In most of the cases, fusion of sperm and egg pronuclei occured outside the body of female, but in rare cases it occured inside the body. The granular cytoplasm displayed a characteristic streaming which leads to pseudo-cleavage after the sperm enn-v but before the fusion of sperm and egg pronuclei. The pseudo-cleavage was followed by the migration of egg pronucleus towards the sperm pronucleus where they oriented themselves slightly obliquely and fused with each other in a plane perpendicular to the longitudinal axis of the egg. Subsequently, the cytoplasm retracted from the shell membrane and the vitelline membrane collapsed as the fertilized egg prepared for cleavage. Pronuclear fusion was completed in 20 to 25 minutes after egg-laying.

Cleavage: The first cleavage was always unequal. occurred 10-15 minutes after the pronuclear fusion, was perpendicular to the longitudinal axis of the egg, and gave rise to a larger anterior blastomere S, and a smaller posterior blastomere P1 . After 5-6 minutes Si divided obliquely into anterior A and posterior B blastomeres. Later Pi also divided into S, and P, giving rise to four-cell stage after 4-5 min. (Fig. 47. B, C).

In the fourth cleavage blastomere .A divided obliquely to the long axis and resulted in

193 the five-cell stage. Shortly. after 4-7 min. the six-cell stage was formed (Fig. 47. D)

by the division of blastomere B. The ten-cell stage and sixteen-cell stage was formed

after 51-61 min. and 68-84 min. respectively, from the first cleavage (Fig. 47. F, H).

Further observations on cleavage became difficult due to the super-imposition of

rapidly dividing cells, constant movement of cells and their dense granulation. The

rapid divisions produced a mass of cells after 70-80 min. after the formation of

sixteen cell-stage, which represented the morula stage (Fig. 47. 1). This was followed

by the blastula stage 90-100 min later (Fig. J). After 80-95 min, gastrula was formed

with two distinct regions, an ectoderm and an endoderm (Fig. K). The cells in one half

which was destined to form the anterior end appeared hyaline while those in the

posterior half showed a dense granular cytoplasm (Fig. 47. L).

After 90-100 min. the Lima bean stage was formed in which the cells on the

ventral side of the embryo started to withdraw inwards to form a notch or depression

(Fig. 48. B). The depression gradually became deeper and formed the Comma stage after 30-40 min (Fig. 48. C). Lima bean and comma stages marked the onset of organogenesis. After 25-35 min. from comma stage. the tadpole stage was formed where a slight depression represented the storraatal cavity (Fig. 48. D). The first twitching movements in the embryo were observed at this stage. The Plum stage formed 30-40 min. later had an elongated narrow posterior part (Fig. 48. E). This stage had a faint stoma outline from the anterior depression and movement became more sustained and smooth. The loop stage was reached subsequently after 40-50 min., where the embryo was about two and a half times of the length of the egg (Fig.

48. F. G). This stage as followed by the early pretzel stage 70-95 min. later. at this stage embryo had a stoma outline, a faint outline of pharyngeo-intestinal junction, lining of rectum and an anal opening demarcating a long, conoid tail posteriorly (Fig.

48. H-L). Intestine and outline of pharynx appeared 30-40 min. later, different parts of

194 the stoma (cheilostom, gymnostom and stegostom) appeared as thin refractory walls.

The early pretzel was followed by the late pretzel 2-3 h later, during this stage the

stoma walls were differentiated into cheilostom, gymnostom and stegostom. Intestine,

pharynx and rectum were well developed (Fig. 48. M, N). Valve plates in the median

bulb were well formed (Fig. 48. M) and the tail narrowed and appeared filiform. The

juvenile attained the length of 3-4 folds and showed the quick movements and even

revolved along its longitudinal axis.

Hatching: After the commencement of the late-pretzel stage, the movement of the

juvenile quickened considerably and occasionally it turned around completely inside

the egg. The length of the juvenile prior to hatching was approximately 3.5-4 egg-

folds. The juvenile exerted a pressure against the egg shell by the repeated labial explorations and thrusting (Fig. 48. P) and by body movements. No moulting of the juvenile was observed before hatching. Prior to hatching, the juvenile frequently forced its lip region against the egg shell. The egg shell became more stretched, elastic and thin. After 3-4 h, the egg shell ruptured and the first-stage juvenile emerged out. It was always the head of the juvenile which emerged out first. Feeding started soon thereafter and particulate matter was seen entering the intestine of the juvenile. The total embryonation time from the single cell egg to hatching was 15-19 h at 30 ± 2"C.

Post-embryogenesis

In studying the post-embryonic development, emphasis was given on the gonad development because the growth patterns of the genital primordium were found to be good markers to differentiate different stages of juvenile.

First stage juveniles: The first stage juveniles measured 168-280 pm in length. A small oval shaped primordium situated at 50 — 57 % of the body length. The

195 primordium measured about 9 — 11 pm in length and 5 — 6 pm in width. In ventral

view the genital primordium appeared slightly obliquely oriented to the main body

axis (Fig. 49. C). In the lateral view it gave the impression of two superimposed

primordial (Fig. 49. A. B). The primordium consisted of two centrally placed

germinal nuclei flanked at anterior and posterior end by two somatic nuclei. About 11

to 26 closely placed ventral chord nuclei (ventral epidermal nuclei) were seen

between the pharynx base and genital primordium (Fig. 49. D). Male and female

juveniles could not be distinguished at this stage of development. During the first

molt, no change in size and shape of primordium was seen.

Second stage juveniles: The second stage juveniles measured about 235 — 404 pm in

length. The primordium elongated slightly without any increase in the number of

cells. The two somatic nuclei became larger (Fig. 49. E-G). The number of ventral

' chord nuclei between the base of pharynx and the genital primordium increased to

about 16 — 28, and were more widely spaced.

The first division within the genital primordium started at the time of second

molting. During the second molt sexual differences in the to sexes became apparent

when the ventral chord nuclei and genital primordium took different courses of

development. In female juveniles the two somatic nuclei divided several times so that

the genital primordium consisted of eight somatic and two germinal nuclei. Two of

the somatic nuclei were cap nuclei. During this molt four to six special ventral chord

nuclei. which were larger and less stained than the ventral chord nuclei, were located

on the ventral side of the primordium. In male juveniles, the anterior somatic nucleus

divided to form six to eight nuclei which later gave rise to male gonoduct. Posterior

nucleus also divided into two nuclei of which one remained as the cap cell and other

migrated to\Nards the ventral side of germinal nuclei. Special ventral chord nuclei were absent in male juveniles. Third stage female juveniles: Third stage juveniles measured 326 — 406 µm in

length. The morphological differences between the two sexes became pronounced in

this stage. The genital primordium increased in length to 20 — 38 µm. The primordium

showed a bidirectional growth and was differentiated into distinct anterior and

posterior lobes (Fig. 50. A-C). The number of germinal nuclei ranged between four to

six which migrated equally into the two developing branches of the primordium. The

somatic nuclei also divided and were equally distributed in both the branches. The

special ventral chord nuclei, the precursors of vulva, were present on the mid-ventral

side of the primordium.

Third stage male juveniles: Third stage male juveniles measured about 273 — 340

inn in length. The length of the primordium ranged between 15 — 22 gm. The

primordium showed anterior growth due to proliferation of the somatic nuclei (Fig.

50. F) and eventually developed a flexure in late third stage (Fig. 50. C. H). The

germinal nuclei and somatic nuclei further increased four to seven and eleven to

fifteen, respectively. The spicular and gubernaculum primordia were represented by

aggregation of dark stained nuclei located on the dorso-lateral side of the rectum (Fig.

50. J).

Fourth stage female juveniles: Fourth stage female juveniles varied in length from

415 — 581 pm. In this stage, there is a considerable elongation in the length of the

gonad. The length of the primordium ranged between 53 — 80 µm. The primordium showed extensive growth anteriorly as well as posteriorly (Fig. 50. D, E). There were

12 — 16 germinal nuclei in each branch, arranged in two rows. Number of special ventral chord nuclei increased to 8 to 10. The specialized ventral chord nuclei aggregated at the site of the vaginal region in a circular manner N%ith an elongated nucleus lying in the centre. The vulvat precursor cells formed the vulva which appeared as a refractive area.

197 During the final moult the primordium increased in length and developed

flexures at the tips. The germinal nuclei were confined to the flexures which formed the ovaries. The cuticular lining of the vagina was also formed during this moult. On completion of the moult the entire reproductive tract was formed.

Fourth stage male juveniles: Fourth stage male juveniles varied in length from 357 —

530 µm. The length of the primordium measured about 47 — 165 µm. The primordium consisted of a short reflexed part containing 7 to 15 germinal nuclei which developed into a testis and a long tubular part having 30 to 75 somatic nuclei which formed the gonoduct in adults (Fig. 50. 1). The germinal nuclei underwent rapid mitotic divisions.

The cap cell remained undivided. The spicular and gubernaculum precursor cells showed more compaction and formed the refractive lining of the spicules and gubernaculum in the late fourth stage, respectively (Fig. 50. K). During the final moulting nine pairs of genital papillae, spicules and gubernaculum developed completely and the gonoduct joined the rectum to form the cloaca.

The total time taken for post-embryonic development was 3 — 4 days.

198 Table 24: Measurements (in um) of different juvenile stages of Myct'olaimus orcodocus sp. n. Mean and S.D. given in parenthesis.

J~ 1: 1, female J) male J, female J, male Adult female Adult male Characters (n=14) (n=14) (n=7) (n=3) (n=8) (n=10) (n=10) (n=14) 168-280 235-404 326 - 406 273-340 415-581 357-530 688-974 623-915 Bali, length 8 (222±35.6) p32.4±39.6) (375.4±27.1) (316.7±30.9) (491 ±53) (447.9±48.9) 785 i 77) (132±50) 10-15 13-18 15-19 13-15 17-23 16-22 Maximum body diam. 25- 30 24- 27 (12.211.6) 115.5} 1.2) (17.3-1.5) (14.3±0.9) 19.611.7) (18.1±2,2) 1.4 20 18.1-228 21-23.7 21-22.7 24-30.6 21.0-31.9 27-32 24.5-29.7 a (18.1 ± 1.3) (21.4 } 1.4) (21.8:0.8) (22.010.7) (25.3±2. (14.9±2.9) (29± 1.4) (27.4:1.4) 2.5-3.3 3.1-4.3 3.3-4.0 3.3-3.6 3.9-5.2 3.4-4.3 4.9-5.8 4.7-5.5 h (2.9}0.3) (3.610.3) (3.8±0.3) (3.4}0.1) (4.3±0.4) (4.0±0.2) (5.3:0.3) ! (5±0.2) 4.1-4.95 4.3-5.5 3.8-5.1 4.5-5.5 4.6-5.7 4.9-14.5 3.9-6 4.6-6.7 C (4.610.2) ! 14.910.31 4.7±0.4) (5.110.4) 5.010.3) (7.312.98) _(j0.6) 5.910.6 44-48 V ------` (46.7f1.2) 33-53 - - (47.815.2) 9-11 10-20 20-38 15-22 53-80 47-165 Primodium length - - (9.910.6) (15.07 2.96) (29.115.9) 1912.9) (71.5.9.2) (83.6±41.3) 5-6 8-9 Primordium«idth 10 7 - - - - (5.6i0.5) (8.5 0.5) 65-84 75-104 95-106 83-1 100-125 104-124 138 -I67 130-175 I'harim (77.2-15.6) (92.717.4) (99.3 3.8) (93.017.5) (115.6 f8) (111.1 ± 6.5) (146-8) (146-12) 90-145 123-196 156-1% 133-170 175-227 164-215 - Primordium from anteriorend (117.17.4) (166117.2) (181.3111.96 (154.3115.6) (207.5116) (195.7± 13.96) 18-13 20-30 25-31 25-29 33-38 30-42 46-56 43-57 Median bulb (length) (2(1.8= 1.4) (25.61 2.7) (28.4-11.8) (26.3 t 1.9) (35.31 1.9) (34.0 ± 3.6) (50±3) (49 ± 3.4) 45-59 55-73 65-75 60-73 73-90 74-87 97-127 99-116 Nerve ring from anterior end 2.S±4.2) (65 f 5.4) (70.7:3.6) (651± 5.6) (82.1 ±4.7) (19.2 f 4.1 ,j5± 8) (105±5) No. of ventral chord nuclei 11-26 16-28 19-26 13-19 19-21 14-24 - _ from basal bulbto rimordium (19,9±4.9) (213:3.1) (2L3*2,3) (16.0±3.0) (2011.0) 11.8±3.3) No. of ventral chord nuclei 37-60 47-67 55-59 52-55 58-63 56-70 _ from basal bulb to anus (48.3± 9.0) (59.8 1 5.6) (57.711.6) (53.5 ± 1.5) (60.5±2.5) (64.315.3) No. of ventral chord nuclei at 3-5 i 4-6 6-10 4-8 12- 12-38 _ primordium (3.8 } 0.8) (4.8 ± 0.8) (1.8 } 1.7) (6±2.0) (i) (17.5: 9.3) - 35-68 50-83 1 75-85 50-75 90 -110 35-90 150-200 110 -145 1'ail (48.4 8.4) (68.3:9.6) ~ (80.4±2.9) (63.3±10.3) (9&6± 6.3) 69.1±20.9) (178±14.9) 126±10) A

B S2 B

S2 P1 P2 P2 A i B C D ►~`'`~ -

E U G

r .

.:.. %- J

Fig. 17. Embryonic stages of alretolaimus orcodoxos sp. n. A. Two cell stage, B. C. Four cell stage, D. Six cell stage. E. Eight cell stage. F. Ten cell stage. G. Twelve cell stage. I-I. sixteen cell stage. I. Morula. J. Blastula, K. Gastrula (early) showing differentiation of ectoderm & endoderm. L. Gastrula (late) showing two regions-h) aline anterior & granular posterior. r ;

B C D __ - j y r. 14

E'... F G H a_

r h l

RL

J K L

M N~ - -- O P i_. -1 S. F mhr\unic sta es of ti. i')luinrtt. or&'o / IVus ,p. n. :1. I,ima bean ,tase t\entral). B. Lima bean stage (lateral). C. Comma stage, I). Iadpole stage, arrow ,ho%Ong the anterior depression, I.. Plum stage. F. G. Loop stage. {I-I.. I:arl pretzel stage. If. arrow showing lining of rectum and anal opening. I. arrow showing outline of stoma. K. arrow showing outline of intestine, L. arrow showing outline of pharvngeo-intestinal junction. M. N. Late pretzel stage. M. arrow showing median bulb with well developed valve plates, N. showing well developed intestine & pharsngeo-intestinal junction, 0. arrow showing genital primordium, P. Labial thrust before hatching. 1 0

Fig 49. Post-embryonic stages of Aftrzuluimus oreodo.rus sp. n. A. B. J (lateral). C. J I (ventral), I). Ventral chord nuclei of JI (ventral), I -G. J2, E. ventral, G. lateral. B

0

t.

F

J

Fig. 50. Post-embryonic stages of AI►•dolaimus ureodoxus sp. n. A. C. J3 female (ventral), arrow showing special ventral chord nuclei. B. J3 female (lateral). D, E. J4 female. F. J3 male (early ), G. 11. J3 male (late). G. ventrally, I1. laterally. I. J4 male, J. J3 male tail, arrow showing spicular & gubernaculum primordia, K. J4 male tail (late), arrow showing the refractive lining of spicules and gubernaculum.

The pattern of cleavage during the embryonic development of Myctolaimus

oreodoxus sp. n. resembles with Cylindrocorpus longistoma described by Chin, 1977.

The first division in the egg is always transverse giving rise to two unequal

blastonteres. Unequal division of blastomeres has also been reported for Rhabdilis

reres by Chuang (1962), Acrobeles complexes by Thomas (1965), Cylindrocorpus

long/stoma by Chin (1977), Oschelus sharnimi by Tahseen & Nisa (2006),

Paroigolaimella bernensis & Fictor anchicoprophaga by Pillai & Taylor (1968).

Anterior blastomere was always found to be larger resembling Caenorhabditic

elegans, Teratorhabditis andrassyi (Tahseen & Jairajpuri, 1988) in contrast to

Ditylenchus triformis (Hirschman, 1962), Ilefieoylenchus vulgaris (Yuen, 1965), If. dihystera ( Hirschnvann & Triantaphyllou, 1967) and Cylindrocorpu.r longiv oma

(Chin, 1977) which possess smaller anterior blastomeres. Cleavage pattern upto four- cell stage ressembled C. elegans ( Von Ehrenstein & Schienerberg, 1980),

Teratorhabditis andrassyi (Tahseen & jairajpuri, 1988), Mononchoides fartideru

(Tahseen et al., 1990), Cglindrocorpus longiswma (Chin, 1977) and Paroigolaimella bernensis & Ficlor anchicoprophaga (Pillai & Taylor, 1968).

After fertilization, the point of sperm entry marked the posterior pole of the egg. Cytoplasmic streaming was also observed in Myctolaimus oreodoxus sp. n leading to pseudocleavage alter sperm entry. Tahscen & Nisa (2006) reported similar occurance in O. shanaimi. During pronuclei fusion, it has been observed that the egg pronueleus moves towards sperm pronuclei and their fusion occurs at the posterior pole of the egg. However, in Teratorhnhditis andrassvi ( Tahseen & jairajpuri, 1988) sperm pronucieus moves towards egg pronucleus and then both the pronuclei shows migration towards the centre which is followed by their fusion but in Chiloplacus symmetricus (Ahmad & jairajpuri, 1977) both the nuclei move towards the centre where they fuse with other.

204 M. oreodoxus sp. n. has a short life cycle as that of other diplogastrid

nematodes. Eggs of M. oreodoxus sp, n. were elongated, smooth-shelled like that of

Mononchoides changi (Hechler, 1970), Mononchoides forlidens (Tahseen el al.,

1990) and Oscheius shamimi (Tahseen & Nisa, 2006). The first polar body was

observed outside the vitelline membrane and second polar body was observed inside

the vitelline membrane which was also observed in Radophofus similis (Van Weerdt,

1960). Rhabditis Ceres (Chuang, 1962) and C. longistoma (Chin, 1977). However, in

Acrebeles complexus (Thomas, 1965) both the polar bodies were observed outside

vitelline tnentbrane.

The intra-uterine development was not observed in M Oreodoxus sp. n. which

has been reported in Mononchoides changi (Hechler, 1970). The presence of only two eggs in the reproductive tract of gravid females of Al. oreodoxus sp. n, suggested late oogenesis as in Mononchoides fortidens (Tahseen et al., 1990) which is in contrast to

Oscheius shamimi (Tahseen & Nisa, 2006) where uterine tract was observed to accommodate as many as fifty eggs. The movement in the embryo for the first time was observed in tadpole stage which is similar to 7eratorhabditis synpapillata

(Tahseen and Jairajpuri, 1988). Oscheius shamimi (Tahseen & Nisa, 2006) and

Mononchoides fortidens (Tahseen er al., 1990). In a few eggs. development of anterior half was arrested due to some unknown reasons which resulted in an embryo with a well developed posterior half and an undifferentiated mass of cells in the anterior half. The posterior half showed vigorous movements for less than 24 hours and then the embryo died before 24 hours.

Hatching occurred as a result of pressure on the egg shell due to growth of the embryo and also due to labial probing against egg shell. These probings were not specifically localized and no blister formation occurred as in case of C. longisfoma

(Chin, 1977). Thus hatching resembles Chiloplacus synmtetrieus (Ahmad & jaimjpuri,

205 1977) and Oscheius shamimi (Tahseen & Nisa, 2006). The head of the juveniles

usually emerged first during eclosion as in case of Paroigolaimella bernensis &

Fictor anchicoprophaga (Pillai & Taylor, 1968). However, Silverman & Campbell

(1959) observed that the juvenile of Haemonchus contortus pierced the egg shell with

its pointed tail and emerged tail first while in A. complexes (Thomas, 1965) the larva

emerged head or tail first depending on its position inside the egg when the membrane

ruptured.

The reproductive system in M. oreodoxus sp. n. was found to develop from a

single obliquely oriented primordium which has also been reported for Mononchoides

fortidens (Tahseen el al., 1990), Diploscapter coronala (Hechler, 1968), D. orientalis

(Tahseen et al., 1990) and Oscheius shamimi (Tahseen & Nisa, 2006). The sexes in

M. oreodoxus sp. n. could be differentiated from the second moult onwards, by the

appearance of specialized ventral chord nuclei opposite to the genital primordium in

females and by differentiated development of the genital primordium in both sexes.

Similar observations were found by Chin (1977) and Tahseen & Nisa (2006) in

Cylindrocorpus Iongistonra and Oscheius shamimi respectively.

The juveniles could not be differentiated exclusively on morphornetric values due to overlap in ranges. But growth patterns of the genital primordium were found to be good markers to differentiate different juvenile stages which has also been reported in Oscheius shamimi (Tahseen & Nisa, 2006). However, it is in contrast to

Cylindrocorpus longistoma (Chin, 1977) where the body length and primordium length of different stages did not overlap. In the first stage juvenile two germinal nuclei were found as in other didelphic species e.g., Helicotylenchus dihystera

(Hirschmann & Triantaphyllou, 1967), Radopholus similis (Van Weerdt, 1960). C. elegans (Von Ehrenstein & Schienerberg, 1980). Mononchoides fortidens (Tahseen et al., 1990). C. longistoma (Chin, 1977) and O. shamimi (Tahseen & Nisa, 2006). In Al. oreodoxus sp. n. it was observed that the number of germinal nuclei remains

unchanged in second stage which has also been reported for Ditylenchus dipsaci

(Yuksel, 1960), D. triformis (Hirschmann, 1962), Helicotylenchus dihystera

(Hirschmann & Triantaphyllou, 1967) and H. vulgari.s (Yuen, 1965), C. longistoma

(Chin, 1977) and Chiloplacus sym netricus (Ahmad & Jairajpuri, 1988). However, it

is in contrast to T. andrassyi (Tahseen & jairajpuri, 1988) which showed a

multiplication of germinal nuclei during the first moulting.

The ontogeny of reproductive system revealed that a continuous and gradual

multiplication of the nuclei in genital primordium occurred throughout the moults and stages as in case of Ditylenchus triformis (Hirschmann, 1962), Pralylenchus sp.

(Roman & Hirschmann, 1969), C. longistoma (Chin, 1977). 0. shamimi (Tahseen &

Nisa, 2006) and Mononchoides fortidens (Tahscen et al., 1990). Therefore, the number of nuclei in the genital primordium at any given stage of development would be subject to variation. However, this is in contrast to reports on H. dihystera

(Hirschmann & Triantaphyllou, 1967) and If vulgaris (Yuen, 1965) where cell multiplication in the genital primordium takes place only during moulting. hence, the number of cells in genital primordium of each developmental stage remains constant.

The primordium of male juveniles exhibited typical secernentean characters which involved elongation of primordium anteriorly. Later, the development of a flexure which grew posteriorly to form a gonoduct was exclusively reported in rhabditids. In females, the flexures of the primordium developed during late fourth stage and contained the germinal nuclei. The special ventral chord nuclei became arranged in a circle which marked the boundary of the vagina. Similar occurrence has been observed in Helicotvlenchus dihystera (I lirschmann & l riantaph} llou. 1967). C. longistoma (Chin, 1977). Al. fortidens (Tahseen et al., 1990) and 0. .shaminii

(Tahseen & Nisa, 2006). The formation of spicules from spicular primordium was

207 morphologically similar to that observed in M. fortidens (Tahseen etal., 1990) and 0. shamimi (Tahseen & Nisa, 2006).

208 S"W" The present work represents a taxonomic study of the nematodes belonging to

suborder Diplogastrina of the order Rhabditida and biological study of a new species

Myetolaimus oreodoxus sp. n. Samples rich in organic matter were collected from

different parts of the country, especially from the northern and north-eastern regions

(Jammu and Kashmir, Uttar Pradesh, Uttarakhand and Manipur). The samples were

collected in different seasons from different types of ecosystems like, farmyard

manure, chicken manure, canals, ditches, ponds, tree-holes, mountains, hills, lakes,

rivers, sewage, leaf-litter, forests, etc. The samples were processed by Cobb's sieving

and decantation and modified Baermann's funnel techniques. The extracted nematode

samples were examined under stereoscopic microscope. Nematodes were

simultaneously killed and fixed in hot F.G. (8 ml of 40% commercial formaldehyde 1-

2 ml of glycerol + 90 ml of distilled water). Nematodes were then transferred to

glycerine-alcohol (5 parts: 95 parts) and were kept in a desiccators for dehydration.

Dehydrated nematodes were mounted in anhydrous glycerine on glass slides using

wax as sealing material. Al! measurements were made on specimens mounted in

dehydrated glycerine with an ocular micrometer. De Man's (1884) formula for

denoting dimensions of nematode was used. Morphological observations and

drawings were made on Olympus BX50 DIC microscope and photographs were made

on Nikon 80i DIC microscope.

In all, twenty-three species belonging to fourteen different genera falling

under seven subfamilies under five different families under two superfamilies of the

suborder Diplogastrina have been described. Of these, fourteen species and one genus

which are new to science and nine already known species have been described and

illustrated in detail. Five of the total fourteen genera and four of nine already known

species have been reported for the first time from India. The embryonic and post- embryonic development of Myctolai.nus oreodoxus sp. n. has also been studied in

209 detail. The process of fertilization, egg-laying, cleavage pattern, activities of first

stage juveniles within the egg, hatching, moulting and copulation were also observed

and studied. The terminology used in the text to describe the parts of stoma is of De

Ley et c11. (1995) and of other structural details of Furst von Lieven and Sudhaus

(2000).

The systematic position of genera and species described in present study are

given below:

Order: Rhabditida

Suborder: Diplogastrina

Superfamilies

1. Diplogasteroidea

2. CyIidrocorporoidea

Families

1. Diplogasteroididae

2. Diplogastridae

3. Neodiplogastridae

4. Ty lopharyngidac

5. Cylindrocorporidae

Subfamilies

1. Rhabditolaiminae

2. Diplogasteroidinae

3. Demaniellinae

4. Diplogastrinae

5. Neodiplogastrinae

210 6. Tylopharynginae

7. Cylindrcorporinae

Genera

1. Goffarlia

2. Rhabditidoides

3. Acrostichus

4. Butlerius

5. Diplogastrellus

6. Pseudacrostichus

7. Demaniella

8. Fictor

9. Koerneria

10. Mononchoides

11. Oigolaimella

12. Pristionchus

13. Ty/ophan'nx

14. Myctolaimus

New genus

1. Pseudacrostichus gen. n.

New Species

1. Goffartia baldus sp. n.

2. Diplogastrellus cun'atus sp. n.

3. Diplogastrellus neograciloides sp. n.

4. Diplogastrellus kashmeriensis sp. n.

5. Diplogastrellus loktakai sp. n.

211 6. Pseudacrostichus musai sp. n.

7. Fictor suptilis sp. n.

8. Fictor longicauda sp. n.

9. Mononchoides tenuispicula sp. n.

10. Mononchoides latigubernacula sp. n.

11. Oigolaimella brevigubernacula sp. n.

12. Pristionchus stercoraris sp. n.

13. Myclolaimus papillatu.s sp. n.

14. Mvctolaimus oreodoxus sp. n

Known species

1. Rhabditidoides papillatus

2. Demaniella keirakensis

3. Acrostichus nudicapitatus

4. Butlerius micans

5. Diplogastrellus grad/is

6. Koerneria frlicaudata

7. Afononchoides striatus

8. Oigolaimella longicauda

9. Tylopharyn x foetida

Genera reported for the first time from India

1. Goffurtia

2. : l cro.stic hu.c

3. Butlerius

4. Pristionchus

5. TvlophalynY

212 Species reported for the first time from India

1. Acrostichus nuducapitatus

2. Butlerius micans

3. Mononchoides striatus

4. Tylopharynx foetida

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