TAXONOMY AND BIONOMICS OF THE
NEMATODE GENUS Butleriuls
CHARLES CLAYTON RUSSELL
A DISSERTATION PRESENTED TO THE GRADUATE COUNCIL OF
THE UNIVERSITY OF FLORIDA
IN PARTIAL FULFILLMlENT OF THE REQUIREMENTS FOR THE
DEGREE OFr DOCTOR OF PHILOSOPHY
UNIVERSITY OF FLORIDA
The wr;-iter wlshe~s ~iatoexrj3ss ;;is apip;`eciat~iT, n I nec ste
D,. V. G. Pe~rr, Chairman, Dr. J. T. Creighton, ?rofessor R. D. cey
Dr. 3, A. Rloberts and yr. G-. C. Smal-t for -their a~ssistance In -;,.
Tp~rhparatin of thsmns7p;t r G. Perry especally for his
ivlu,~Iable sugge,7stions t, `Ou';3;, Tc couragement am critiism trouhus 'Ch
wrtr' gIaduae studyr; to ?rfsc:.T C'wyfo rviino
facilities an~d financial assis-,ance thr;;ugh ;:atio-nal Insti,ute ol
Health ~ OC~O grn .P 00;to 7~. n. rA. 0Nove and 3r E. ~1. Mille, for
,ssis-tance in- the preproduction of the ?lates; an d finally,, to his kif
for her unide~atancng and assist~ar e caring this pe~riod of gradute
~c knowledge Its .......,,.......... .................. ...........
List of Taoles . . . . . . . . . . . . .
List of Platres ..................................... .... ...
List outof, Fig es..........................................
Materials and ;lethods
G-en~eral Techniques ........
IFsr he ogy and Taxtonomy ..
Results and Discussion
End~l `l 12ci~ sj .. ............... .....
Desc;igioln ................... ......
Diagnosis .... . .. . .. ..
Holot p~e .............................
Par8 OS7 .............*************-*
Biione c.Ii ............................
ne .a ..........................
O 12;ir g ......*******************
Re -oduction ....................
eed~ sru ....................
ie~ sin ..........................~
Summ~ary .................. ................... ................. 75
Literature Cited ........................................... 77
Biographical Sketch ......................................... 79
^.n: oduction .........
Pu a a~3. as u....
Bianc as .... .. . .
~ife c-o e.........
Repr oductio n .. . . . .. . . .. .
Table 1. Larval St73e Duration and Body Sengths
of En Ia.-rlus 1-Ci .s ...,..
Table 2. Larva;l Stage Dur~ation and Boacy Leng chs
Table 3. Larva; Stage Duration and Body Lan ths
~table 4. Average Time Lapse of the Developaunal Sta es
of Bujtlerius spp. (in, bra r'rom ovipositin- a-,
27 . . .. . .. .. . .. .. .. . 75
IST ,F P ATESj
J..ktO I. E8_l- it :--S r. Sp. ................... .......... 23
?late II. Eutilerils r- -slus n. sp. ................... ....... C2
Ilate III. 3 l~ eiv .s.................. ....... 6L
L^S;' uF HOURES~
Zi. mryronic deveLopm~ent of Ir u-~u lac~rs ............. 28
Fig 2. Ccpulation~r of -vie -i lh ............
Fig. 3. Fecing of _periu l.aci~ .......................... 37
Fig. I;. Dmor~ryon~ic dcvelvupment of iRlru e
to the morula stage ............................... 4b
"i 5. Embryonic developmrernt of E-t-eri s rssalas
fr"omr blastula to eclosion ......................... 48
?i.7. Rlting of L t a snoa ......................... 5%
Fig,6 6. Copulation of Buti T us~ ra.;. 1 ...................... i7
Fig. 9. Feeding of Sui~t 5 ? sal ......................... :9
FIg. 10. Em,~bron;ic cavelopmentt ofr F e.6J 'ar .:so ce .......... 68
Fig.11. Copulation, o; B s .................. .. 7
Fig. 12. Cannibalism by F. -is -2. aricas ................... 7~
The gen~us En;tleeius T. Goodey, -929, was erected as a monospecific
ge:;us with the type; sFcies, "L. EU =ri, nBd after' Cz. 3. J. Butler,
the collector. T:,e descr-iption wls cased on nirne specimens extracted
f'romn rotted banana roots prcserved in "strong~ alcohol.' The stomnal
mrorpolology of B ariusT'; `r--" w~as originally desci:il 1 oy T. Doc-'ey
(5) as follows:
The head is ,:.,;r croadiL~l -and ?nreiry lteed
center It is raised no, In the lorm~ of` a ';cruncated con~e wlth The
mouth apertulre in fro~nt. There ca liot' ap-ear to bc Eany distinct
lips a-d th;e walls of' th~e conical region seelr ra,her thin and
membpranous :. Thre are sixe ceph~alic pcap~illae equallyr spaced -round
the head e:,d behind the: sor~ical :much rtegion. isach has a fai-lry
wide basal Dat L hl~ ch ca;-ies a mroc-ratelyr long sets. Ampair~ds
wjere noct observed;.
As seen in optical sec-,ion (fi'. 20), tae; cucal capsule is
mae p fL'o priinc;i Dal ~ part;. an anterior 'ouccal arnd a posteror
;har~yngeal region a atlieo erctnbeeethto
oeing seen runnclr across the capstle. Tee focr:-.rd part of the
-bucc~ ` alls f'orm une t-runcate~d coc~a of' thl ;uuth. They are
relatively thin and byallie but gradually inre9se in thickness
pcstoriorly~ where they art~iculate :.;th the pha~rynga~l pa-rt. The
la~tter is in the f'orrm of a h~ollowJ ring w~ith? con~vex wa:lls w;;ich
are thicker posteriorly tha~n ante;-~riorly The base of h iio
each size is expanded and is attached to the 3-nterior elcd of the
oesophagus where it curves outua~rd. The dorsal side Is a li-tle
In &avance cf th~e ven-tral. ~The cuticle coverl;,i'ng th aerior and
of th-e cesoph~agus is relatively t;;ick and is prc;cineed into tw~o
inw~ardljy pointing teeth; one dorsal and the other vcntral, the
former a little in front of' the latter. These twJo teeth ae:
fojnd on either side of an almost cylindrical hollow let into -,he
anterior face of the oesophagus, a ,Litle d~eeper thran w~ide and
having the lumen of" tae oesophagus leading out of its 1cor.
Goodey further stated.
Another dIffere:ne shownUr by 3 21 tl1-'S is in the bu~ccai
cap~sule, the grea-t size and structures of a-nich sepa~ate it frc:?
nlo-s e Th two; "fixd teeth gua-rding the- ent.-rance to the
hlollow. cavity in? the anteirior e,,d of t:-e oe;ophagus ar-e agair
different I;om tne noviole teethr fon nsm eiso
?'i-i- tor Te ccav;ityr ;ist men~tioned Is an~octher distinctive
feature of th-e newi gert~s. Th ltarei edwhtemuh
on the summit of tha.) uru:: i~' eaed r.-, the sides of wir:.ch: are with-;-
cut radlially sei;-eami~ loo~f-crownT elements an! the absen~ce of _ips
m~arkl it off` fr-om ;C.? it-
The females of' this species were described as possess..; a vulva
at 58 to 64- per cen~t didelphric reflexed ovaries. h.nehaa
single reflexed testis well-d~~eveled spclules ,.,, Lzccoring
Goodey, "a remarkabl- --:ge gueCcl.. ooday furier stated-:
"TheL gu'oernTaculum is more than, ta.c-thirds tlhu length~- o' -,ne 3picue;
its head and lies close to -,hce dorsal s dS of tne bodJ werll ard is
curved inwrd;;rs ,wa~rds the~ socules in~ a snall exuar,~sion o_ ,ockc. Th
cody of the g-jbelr;,aculum~, is hoilc=; and ir. ,ho distal ..1:& of its
lenthitseemrs to encos th aei; iuLes which~., pass complete
thrown it O ach side: of its ventral er., there Is a s.-.l; latera~l
E cG m ent io n h~a s m ad e of- t o i e b t r : o d h o it s a
In; 1930, InuamT (:) d~ebCozibed r. 1 isdat f-.. sp3eime;-s
extracted from a com.post heapj. AOam'sS Cdesriptic.. wras Bas~e on bojtr
l'iviTn and; heat-r-elaxed specime2na .cst~ L ;;im: nc.c::ni;.
.no-1p-ological character sxmvai lp acconLi 1.e
sulrface with the anteriorrost nort-ion~f of ,;;3 chaliorh;dir.,the3ip
sliightly overlap arnd unite to form a somewha'T, fLattr cr; si
la'oial papllza each vweari -ng o unequal~ setae or crstl :. flxibl
r,:n, connecti~ng the~ aneiradpsarl~ po`tf9ons of the .;tor.,;
1,rge corsal tooth whiah i peroraew acn1 nlnly
a'; -3 ar cent and -the ,-istll tends of the two~ re-"lexed- o-.i.ie, externded
back~ to th-e vjlva. T,1e testis wras reflex-ed and si;-ply constructfed.
Spicu~les we-re hav~.IJ sElecti ad1 and. elong te~d.
I el~iev.e, after studyingl my living~ material as c os-ely as
possible, that a more precise study of' Goodey's material would
1. Tae ocene described by Goodey in B. ;~tle-i consisted
of six movaole lips. Therefore e lips are not missing as
he indicates In his descriotic
2. The cephalic s .millme: of 3. t;.r'er perhaps conii~st
of" two se:,ie a;Td nojt on~.3
3.The t, .u chit inied s-;omal par-ts are separated by an
expanding memoranouss rirg, and not, 7 C-ooday said, ty meals~
of a 'faint ,1 a of dtmarcation.' The latze: is probably no
lor.,: to be seen on fixed ma.terial as it is severely con-
tr-acted on, thre ducih of th~e animal.
F~inllly t.:ere are p-cobily two ven~tral ttethr in the
case o;' B. `Dut i also as they; scarcely can De seen wthl~`out
1.-38, B. exe and -. `r isnicu~llllus Schut~ mans-Stikhoven~ and
Tennissen wre~e described. The cascription of B. okai (11) was cased on
taizzycl~-nince specimens extrac~ted fromn soil moss In Hainan. According to
RE:, ; precise descrip'ion- w~as not possible blcausee of (translation
frcm Ge~rman IIpost-mo;ryem alterai-on (particularly in? the position of
,,.e sevxu~al o",ans)."
Rahm describes four teeth in the stoma of E. 2 1_ ; the two~ sub-
ven-,rals as described by Aiam~, a larde dorsal and a smaller dorsal
tooth, apparent.cly in tandem with the larger one. He was un~acle to
corrobor;ae eithu: toe canal in the dorsal tooth or the anterio-
;'chitiniz7ed ring," dscrib~ed by Ada:-. R~ahm, stted! that he was unt;le
_-a 3;, ;Ixiopt r'r the. v.1va whichh wra; at 4S to 51 p~e' cei..) d~ taie
hc=vi~ cuticul~a-izedi spicules (not reporced t;o be accompanied by a
Rahm,7 su plied no bionomical inf~ormnatior- pertaining to B. C'Kr3i
other -,han to h-pothesi e that t~he elastic ri, observed by both Acav.
andhi, might m~ake pssible the functi ~nin or h niesoaa
Schuinarmas-Ste xhoven and Te aniss; n 13 ) 2e scribed B. bare~ s~~nir -
vices.~ iro;rs a single male collected rfron a baboo restt In. Lte C; 7c.
They- were un~able to give much detail on the structure of the stomra
because of its state of' cont;ctior.. They did stale, howeve-, that -the
c31inczrical stoma was arm~ed with: two curved pointed Leeth. The robust
heavily scl;rotlized, nioncephalaied! spicule; wJere vquipped with a
d~istirnct long~itudinal ridge stubc~ ex--en~ded =leir en-,ire legth. e
gubjernac;ulum was rcbust and it; lengthlr was one~-half thatc of the sp~icules.
No nentica was male of the bionomics of this ,,,cies.
In hio book;, S--1 a-d -r~eshwatetr :swat-~ls, i'. Ca~cday ;6) agroad
itl-h Adam's inte~rpretation of the lip structure of' B tler U~s and,
basea on stomal morhology, suggested that this genus might have a
IKey1 (9) describied 1. Ferl ti,, using; tw,enty-rtwo female ar.d th~re
ale; sp~cimens collected ;'rom the water reserve cf the bromreliad,
js elia arensis (Vell.) Mez. in Brazil. His desc--iptior. of the
stoma is in emplvte agr~eL-ment with that of' L0 day (i). Th're female
specliness wer~e d~e32coibed as hbving almost syrletrictll amphidelpchic
rifleX;~i CVaries rachnlrg almost halfw~ay to mne vu~lva w~hich I.as locacted
at -,,.2 7;o -7.5 p~er cenL.; The -uticle sras finly~l an~nula -d ndrc .lad
ten lonrgitudical strilae. The spicules were eq~uipped wit;; knobboed heads
follcw~ed by an expanded section- whtic.. taper-ed to sha-~p tips. The
g~~ace:iLaclu has ;ar e, 'olad~;e-lke elaborated proximally, a-d formed
a c~ff~ at th~e cloacul end in;oie of whJIcLh the cLapered ends of the
;e7, listed the focd habits cf B. Earlachi1 as unknowLn, but suggested
that the spcies pobably feeds on protozoa.
"he second Erazllan, species, . FirJ .5 was =oLlected ,,y
Lordello and Zai~th (s) from soil around the roots o' cultivated C :ica
anava~ L. The description of this species was based on both free, and
The descr-iptionr of the: stcoma of sircu~~lr Is similliar to that
of 'c-lei. oc of thre s'jx setose pap:i;E e erm~inate irn a roundec
bodyr of variable s ze. Th~e vulv~ is located at 4iL.3 p~el cent and t:,e
twol -refllax;d o3varies are of unq~ual lenrgth wi~th eithe; ovary bei.g, the
l'onr he s ngle tests of ihe male Is r;flexed; -th-e ccophalated
spicules are strcrg, curved and their tips are surrounded by the
guornaulu. cso--ding to Lorcello and Zamith, th~e most dilstinctive
mo-;phological character of this species is an unpair~ed phasmi~d-like
o-,an for rabich they proposed the cnme parahscd
Lord~ello and Zt~raith sctudied the. focd habi'its of' B. Sirr laris by
rrounrting sp~ci-ens in wtate be-tween a slide and L cover,-lass. iThey
determ~ined that B. singularis was predaceous in habit. They were unable
to recognize th~e vilctims, butl stated t;.at, with little doubt, some of
the. victims were~ worker sp~ecime~s of' E. si-7fe-is.
In te same~ --a-er Lorma~llo a;-d Za.1ith arected to. genus
St~e'kh.ren~ and Teisen to the genu;s 1 Shl :.ar ,
;Adam ;s the type~ spcies. E-;-''-^-1 6 was char~a.-2cteize as hzi
th:e dorsal t-ansition Ec;a ::i-, a 1&: a- hollow co,3l too~:..; teloCs1 C.0
as an oesops;~iagea cy~linde;; six ap c c~~, ^ papi;tae equiipped ;rith 1;;ire
o~ristles w;;ich are appr-oxim~auel- 53 pe; centl of th~e head~ :ri-iat; a..d
six movable, anteriorly prtui. is
in the revised second~ edition~ of $-'1 F re
J. B., Goodey (7) rejected Lordello and ZcUzait-'s t-a-sfer of .'r-1
sniulatus Schuurmnans-Stekh"ovenr and Teun~iasen to PS.- s :c- and -the
erection of Bjutler ? Bes to con~tain .-' :Rahm as w~ell as Meyrl's
t;-rasf'er to ; '. fili-:; 5ts to the n~ewilyr e cted L1 __i _- H
stated thiat, "thre desc-iutions or' E. b- irl eig-::-.;- and E~. ek=:
approchck the cLteg~ory of s--acf.-; rLan-- an ~ljor. c .;S se yundts .
actions of neyl, 1960 andc ;o dello and Za-.it, 1959 a-e not acceptd
On teeoseGocysonr-ized all three~ new: c;:.,Si ain ihL
originaly proposd namesj resu;lcigi ttlo ixseis n 1.
The seventh' species of ^-- `_- :5 rl Jcribed was .
Taylor, 196 1).Tisi a mon;ed-1phicl species -.iT;.. ,.-. ul.:
located at 48 to 50 per cent. '=he testis is, single refllexed, a:- 1-.
splicules are venitrally a~cuate. Ta~ylor de~scr-ibed the '"rncu as
comp.lex~> ad lilclust~rate It .10 Ltaing more- c, sn half as doep: rs -vj.
We ~ ~ ~ ~ ~ ~ ~ ~~L mopolg f h irtto hfero heaea C3~lr3 Sponse
ve~=; clos--ly to th;, deou~~ibac Sb previous au~th~ors. Jampar a .s,~, of' t;;
posterior cham'oer ol this species with others is impossible since
Taylc-'s desc-iption st~ated, "Posterior ch~arrber cen-t it;-s a large dorsal
tocth and other smral`_er teeth anid raispin-g sta-ctures as shownr in Fig.
Taylor's extensive observations of the feeding habtis of E. mo?-
hystera are quoted In thei_ entirety below.
The feeding of B. r: nt -stars has been observed, and .his
species is predaceous upon otner nematodes, as has been reported
by Lordello an-d Zanlith (1959) for B. sicrmis ics. Mos-i specirners
observed attacked&c -Em~atodes havJing :. cony diamier smaller than
-themrselves. The prey was usually inpugzted tail, or less commoni;ly,
head-first. WhJlen a-ttacking its prey, -che p~redator's stoma becaml:e
short~~ene, caused oy a contrac~tion: of the noi--sclera~ti, ed m~iddle
chamber, and -meth? in t::a oasal cham-ber ;; re thu;s able to pEuncture~
the prey's body wall. Body conternts oI the prey we~re i-ge rued and
o~;ved posce-riorly in tie Ulumn of th~e pre~da-or' s procorpus.
Passage~ of food was a~ccomp-anied? by ex~treme d-al~ation of the,~ laz-e~r..
Food -low was less ra idl in ,rhe p~oster-ior portion of t:;e oesopnagus
and was~ _ot Gcompanied by an expansion sf the luner., indicatin
thre proba'le valvular function of the sclerotized plates in tne
metacorp~us. Failure of this species to become established on a
cul;ture ofI '-ele-- b-ur a7;rae Ba;-cian, 1865, prevented additional
TIayrlor s- Dor-,ed LOiey's SynOrLOrz~iZ~ill Of EnTlepei- lS~ fi'ii-
;a ;-s (Ad::, '930) -Xeyl, 196u; 'u';-l-ricide _kai (Rahnm, 1938),
Lordeill am eit,199 a;d ri-.l-raster b-evis i~iclat s ~c:?uurman~s-
Stekhove-Jn andi Tau~nissen,, c938) ;ordello and Zalmithr, 1959. He presented
fathr agumets or the synno~omizatio- oi the latter twjo combiratins
but- like Gooday, piesented no ju~stification for synroronization of
IT t-ell_-s filio=.vit~ tus ( dam, 1930) M~ey~l, 1960. At present tne genus
3. t 1:-1 01 G-oudevy, '9'9; typ~e sp~cies
2.ae~l si~ -1^.- Schcuur::~.a s- Stekh oven an d Teun~i sse n, 1936
C1. D'-_ .r: bl-rev;i~a-l?-ates (Sch;uurmalns-Stekhiove-r and
Counisse, rE 13; ordello are lamith, :959
3. OF -1 Riahm~, 19 8
jyn. Bu~tlerlio5 rs okiai (Rabs?, 1938) Lordello and Zamithr, 19i59
s. sinzLul ris Lordello and Zamit' 1959
It is Lpparent from the descriptions that the above species exhibit
an unusual dcgree of _crphological variation for a genas, 6nd tnat the
inform~ation on their bioanmics is meager at bes,. Therefore, upon the
recovery of three nomatede populations which were identified as unde-
scribed species of the iienus 31ut -rius, a program of research wras
initiated .o (1) prepare ducsciption~s of the nrew~ species and to make
detailed studius of the: (2) embryology; (3) larval development;
(4) molting process; (5) reproductive and (6) feeding habits of each
Fielo samples of varying volumes werre obtained and tran~sported to
the laboratory in plastic bagas; nem;azodes we~re extracted by a miodifi-
cation of the Seinhorst sedimentation process described by G-oodey (4).
Spcim;ens of each species of -1utleries wjere then hna-d-;picked to 1 per
cent water agar In separate Petri dishes, and several hunared specimens
of 3-- rlrlins -re~fT-i ra (;ina., 1767) Good~y, 1965 we~re ;ntroduced
Into each~ dish zj a f~oo sour~e. IAfter the colonies of Eule-i- 1s
jecame-. established, eachl specie, was subscolonirzed byi transferrirg ten
mrales and a single gravid fe,;ale of each species to an agar plate. rAll
indlividualls used in the followringc studies wJere obtain~ed fromn stocked
colon es estaulished from ,he progeny of these females.
Stock colonies were maircained in 9C-mmr Petri dishes flle~d to a
dept ofapproxiately 5 im wital 0.75 per cent water a~gr. Pyxad
polystYrene dishes were used. Th'e dishes we;re labeled with an in~delible
marker on the side of the bot-sm section rather th,, on tihe snace
provided on ,he over. ObS~ervation~s of the near.atodes woire not hampered
by th-e latte in this wayr and~ the colony rer.aineid properly labeled, eve~n
through the cover-s ,Ight have been sw~itched during routil-e colony
Coloni~es uf '.7rcelois-es sp., 3revrib-cca sc., 3:- _:TdTroccousi sp.,
r al' -s -i; vl-l; Pennerr nl~imus sp. anrd RF .bdit's s-pp. were
maintain-ed for use as p3~eyr in s udyinlrg ,if^ hilsto-ries ana Iood backs
Colonies of Par.i"_-:el- re~~l; vi 5 wer esta'ollished or- cooke.~ ,t-
Ireal placed in the centers of 1.5-1 widleccsh fruit Jars and covre=L-c wis
ci,cular panes of" glass, Water the colornies were well1 estao1ished, tie
nematodes were removed from the sides of a jar with a sc=a-re constucted
by inserting half of a dou'ole-edged razor 'olade in c wooden b ,le. .:.e
nemnatodes then were w~arh:ed to a 325-mash~ sc;-,e;. aza ri:C.S, T.t.: runnTi
wrater. The speciners wrere then. -,s;,ed into~ a on et r .:it; a 3orXplus c^
water, rnd allowscc J ;ottle. Then tr,yj c. daw;n i to a 53-.. -1 ;,t:=
fitted; with a rubue; bult cand allowed" to concenrt_,at: in-. ir x
tip~ of thle pipettte. ThafclttdterLa~lr diraty o t
colonies of 5.,3;'_ la13 wJich; a minimumT ofvrr.Te:eerd a
anoC :ettling e-LImInatced most 0:Lebakl c ya~sts tha, would
o'h;erwise co mnatet a;d discolo:~ -,he agT.
The ota~er genera listed Ecoveee ;;r roar~ed c. 1 per" C, wte ai
witai Me:trcal as the sour-ce of 1Iutri et (12). nahg opltc
of nematodes had bceen reac;;ed, -in, surface of the agrwa e~ntly
flooded wijthn water. Firve to1 'nltrte aecnann h
prey~j nem~a-,ones;s was ;`2 poure ont a 325-;:esn s ~c;-ree cT. 'a a on am~C
settling proce~dure outli:1ed fo:` re'O s was f110we
Pr-ey nematdes th;-at had~ belen conen~trated In the; dlroppet~ r w:ere
tran11sferre to a sn;l Syracuse (Buoa of antiiv cr p):&L
glass and w~ere hranid-picked! cith a dent~al pulp canal~ filewh s l
numo -s of preyr were requirell. Stock~ colo'riesj of I cr_ as op
and D -L -i'-s er': ils eemitie n es;ae manner as .L'- 1
colonies. Aill studies were coal~ucted at a rcom: ter-eerature of
M~o ,ihology sr.d "axc,.ca,
"he morphology a~ living specimiens of' -he three u:ndescribed
;secies of r--i-l- in was s-ludied at magnliftlcaton up to 1000x.
Speci .en-s were mounted in as~ albumrEnh, cai comnpress mou;-.-s (descrlibed
beloww, or on ag-ar slide preparations to slowJ dowan -their movements.
C;;mpass mounts were psrepared by placing~ living ne;:atodes in a
drop of water on a clean microscope slide. A glass rod with a diameter
very slightly less than ,;e greatest body ciameter of ,; specimen was
t~ransferred to the d-op. The rod wras broken intco four small pieces and
r~ranged so as to come to rest near the corners of a 24 x 24 mm cover-
glass which w;as then placed on the water d:0~ple~t. The coverglass
floated on -the excess water and coula be cen~terecd on ~.3 slide w:ithcut
rolling the specimens. he~ excess water then was removed and simultare-
ously the speci r s we~re drawnr to t:;e cente-r of the m~ount~ by application.
of fllter paper psointis to appropriate edges of the coverglass. Wh~en
,ee covle-glass had come to res-, on ,he gasss rods, two of its diagonally
cppossing co-ners were fastened. with wax~ droplets fromn a ;rmall candle.
Cal.y -two corners of th-e ccverglass so fastene~d p;-ovedl suff~icien~t to
prevernt rolin, of thne specirzens, yet allowe-id the coveral
By ;-is technclce, the observer was able to control the amnoun, c_
pressure brought to bear c_ the specime-s by adldition or withd~arawl of
water f;rom urser the coverglass; thus, control was maintained over the
degree of freedom of movement allowed the specimen. Staining in viv'
was greatly facilitated because dilute stains could be added to th~e
eage of the coverglass.
Agar slide mounts -.ere prepared 'oy stirring autoclaved 2 per cent
water a ar wri-h L. ma rnamic s-ii-rrr until a temp-r.Laure o- LL C ':a1s
resched. A d;`cp of' c1ar was ti.,n LLrlnfr ed~ to th~re Surfa~ce3 of a ~-Cluer
microscope slide anc quickly spread t;o ; thicl.-e s of Lpproxinately
1 mm1. After the agar had solidified, the specimens to be studied w;ere
transferred to a ey ml drojlet of wtei- ;n the surface of trot agar
and a 21e x 4C8 mmi coverglass was applie,. The mo~u:.t wa~s then. schl-d
comp~letely with waxz~ fr~om : small candle~. Th;e 5inal aga: cornctntrationr
obtained by this method w~as su::.ewhat- g~reater -thanr the ir.itia 2 perL cernt
and the nemnatodes' m~ovtemen-,s w~e~ csnsiderably i..:eded=, thus, a''for"ig
observations into the bodies of the sclrecians from. all C1irctions.
Several accepted methods for killing-. r~e:atedes for microscopic
examination were uoed, but the, t~echnique dr ribed below~ proveos tae
mrost satisfactory in redu;cin p, ost-mortem distc;-t-on. S~ci:s four
;lhc stock colonies werer waEs;heI byr picklng -them fiist ;r.vo a -:a=:: ; coo3
containing water and~ thr..n In~to a 35-;-:r. polyoye:. Peri dish :a '-filled~
with water. The Petri di~~sh ws thn r'loa'Ted In d csoverea in~suated
contiainer th~ree-fo urths filled w~ithr nt:2r id at u ti;.per,2tlre
5c000 and sti;rree slowliy on a magnetic stir"e, La maintain can Lant
wate; tempieratLre t;hroughout th're bath. The~ dish -rlremind In -,he: cash~
for 10 min, wa~s theni rem7oved a;-dC tl;e remaitodes~ .;ere curredtc cr a c-ean
g~iass slide following scandari techniques for ->,oc wtr..L.a()
Specimenls to bc jtual~jed after~1 _ixutlio or pr cese -o lycoix
fo~r permanent mnouns wer roae yth bv eho;E-:o ~:se
to the hot water bath, a eod3-a plsyeePt dlSh conz C1;-Iri
a quantity of TAIF fixative equal to t:;e; quantity' 3 of w e" i' Ln te is
dish also w~as placed i;, tae bath. Atr1 itefxtv a m
to the dish which contain~red the nemnatsc:s. Th~e dishr wc~as ;ihe: placicd cn
a oisture chavi.'er 'or a miiu of 2 br o iztc.Colwn
rixatlion, tihe speeln~rs 5,are ,counTied -,a: or,=il, In T..F3 oolu~t-un~ crr.
st-udied or allowed to remain in thre .iois,-,re ch,;e-te un~til needs; for
Sp~ecimens to 'oe permanenertly mounted we;re fixol, fr' a least 2i hra
and then processed! to desiccated glycerin-e following, the Seirnholt mretho-d
(4.Specimren3 were then mounted in desi-cated glycerin~e between tw~o
coverglasses on Cobb alumin;uml slides folicaring the methods o" Thorne (15),
Desc;iptions~ were prepa~,ed usingL the m~orph-olo;Lcal terminolog.l~y of
Chitw~Nood (3). Ilusrtin were prepared using the met-ods described
by Thorne (15). Photomicrographic techniques were also dzvelope, Ior
Photomlcrob-raphs? of fixed an pemnnLy .:uted camatedes wuitre
made on 35-mmi~ Kodak Fine C--ain Positive "'.. Hger zqality prlnts
rilns used. C'oser-vation of film du;-rin cevlopm19.~ eI b,; use Of a safe--
light made it possi'cle to ctr eloenof oh 2,6~" neai o t..at
th-e desired structuires ccold be accen-,t;Ca. -ine iLevin ?csi-,ive ij
aslow~ film (ASA 3) thtr'ie xoue fu to 4J sec. It cnno
be used, -cherefo;-e, on;liv~ng ;material or in lS~iaboratoric i.,ere micrc-
scope vibration occurs.
al; eggs weare obta2ired fo we. ,rs-fe omso o.Goft0
g-ravi; fema~les w~ere t;-ransftered f~rom ,cha colony, to watch. glas
containing deio.;ized water, Ley w :re tn trn2rras d~crle,
of' deionizer; wa-,er in the center of a 19 x 24 r~n sc1:i__e cLverglass~~
whilchc was then inverted on a hang~ir. drcp, microculture slide -;-:itcou;
ringSng. The f~emalfs were obse~.ved -:ith -1-3 a1d c .Isscti-?g c.ic:
scopTe until three or more- Eggs,:, `5E~ bee1i:; it his',1 timeC th~e 3ove.--
gla~ss waJ~s tur ned ove;, the feimales 1e 3va D S poi3ioned in?
the center of the d;-oplet f"ree fromn surfa-ce tension of' the waste,. n.
droplet of 2 per cent wJater agar that~ ihad been~- cooled with agitationn
(37 C) was quickly added to the water droplet. e h arha
solidified, thre coverglass was inverted on~to a hungr.ir.; drop ;r.icr~culth~; e
slide, previously ring;ed wilth petro;~Lum jelly, so that the ager Crop
was placed in the polished 13 x( 1.?5 rrm straignt-wdalksG COr~cavly. .
coverglass was then depressed a-d saignaly rotamed ;-cutil a iac.,pi;-,; ;al
was obtained. The sde wa~s co;;sta~tly otum ved co outh :op a
up to 1000x at linervals dictated by the degree of cha go ooserved ,.-
the e gs. The lapsed timre was rezor-ded f'o- eac,; photc.:icrograph.?
On t tree slnil arlyr prepa re l- 2 d o l k ~ c n u n n
from tinree to ten egg-s of on~e ;c.;atode sF'cieweeabave t af
hour, inte;vals through~out development.-~ Th~ oaser;-vatic.-s Twere ac.;dutec
using a separatep researchi ~.l.~croscpe ajaent, to ta;e oeue o
compilete embr~ryology stazy and p~hctLomi,croE;-.: phic series. Stage i
development at each~ obser-vation; period wasreco6- l'3CtCu Iv" e se-o.;d
series of egg,o, but no ph'otomricrographsi were takten.
Prelimin;rarj stludies of larval, deralopmentr~ werec-dledb
c senvrvin the jevelopme;-t of indij~viduu1 larvae th~in t..ic lawyers ci
water aga-r held iin polyrstyrene Fet-ri dishes of' either a 90-mm, 55-;n:<,
or 35-m~a size. ,se of the two lar,,r dishes proved impr-acticable as
excessive time wras required to locatt active But~lerie larvae following
egress from the eggs. The 35-nmm dishes proved unsatisfaoctry because
the thin lawyer of agar dried cut before the nematode reached the adult
stage. Th-e techn~icue described below was developed to allow~ observation
of large numbers of developing larvae and minimize extpenditure of time
and agar desizcation.
Petri dishes of a 90-mm,7 ,ia ete-r we-re 'illed with 3.75 per cen~t
water aga~r of' approxcima~tely 2 mmn by swirlingr, the agar in the dish until
It had becomTe viscous. After the 2agar ,ad solidified, seven disks,
20 ,7mr in asiameter, were cut with a cork borer. One disk was cuti in -the
center of th~e ;late an~d the other six were spaced ecuidlstan~t around
th-e center. The plate was then inverted and the agar between the dissks
w~as rmoved and discarded~. A- single e g of Butle-iuls in a larval st~age
of cavlcpm~ent was t;-rasferr~ea rom the stock: colony to the center of
each agar dissk. Specimens of Zana~rrellus radivir ers~ were inr~;oduced,,
and each disk was covered with a diskc which previously had been cut
with a sharp cork borer from a th~in plastic fIle. "he plates were then
Inv~erted onto t:1e stage of' a research m;icroscopje and observed at
.iagnifications us to 200x. The larvrae were observed t 2- to G-br
intervals throu~ghouL~ development. Between observations th~e plates
were ktept in inverted metal 10-cm diameter cans equipped writh plastic
35elf-sealing lis these served as moisture chambers.
although rColting was 3obse-_ved inl ail la1rvl stages, dlevailed
;tuu~ies here co..ducti-d on freshlyr fed 'ourth-ssaage larvae. Four-th-
stage larvae were selected? fol st-idy because they irnitiated molting
soon after feeding, and thair larger size f'acliltated detailed
observations of rhabdional migration and formation.
'Two techniques were used to study the molting process. In the
first, a thini plastic film was placed in the i:;vert~ed cover of a 90-mm~r
polystyren~e Petri dish; a small quantity of 1 per cent water agar w~as
poured onto the plastic film. The bottom of the Petri dish wass pressed
into the cover and held 'oy pressure u:-til the agar had solidified. The
cover was the;; removed leaving a 1-mm? layer of crater aga- between. the
dish bottom arno th-e plastic film. A small "V" w~as cut in the plastic
film and the apox puclled back to expose a section ;f toe agar. Three to
five 13Vlart wGere t;-an-sferred to th-e exosed aga;- surface and! the plastic
was replaced. The molting process wus observed through~ the plajtic film
wi~ta the aid of a microscope equipped with an oil-icmmersion objctive.
This technique allowed ample oxygen to reach the agar throLt the
plastic film and -the low prof3;ile of t;he mount allowed easy, access to thei
The second technique, used only forobingphticgrhs
involved the use of the~ agar slide preparazionss described above.
Spcien o revius 3. w'ere frequentlyj observed in. em-ult:riu
in stc=;e colonies, a,; more detalet studiess were made of virgin laemales
oat..;.ea fromr larva1ldevelopment stuaies or other individuals espec~illy
roarted from; larvae-. Males that had not rated! wIere obtain~ed in the sam~e
mrann~er as the fealle.; males that pres~nmably had mated .:ere obtained
fro- the stock colonies. Detailed cbjaervatirns were made at r:.;gni'ication
upJ to 1000x by acth r~chr.iq~ues desc-ibed above under "M~olting." Observa-
tions of behavorial resccnses t- males to the introduction of virgin
females into heavily p3opulazed szockl colonies and res onsjes to single
virgin femhles in agar plates were made as magnifizaticas up to 20Cx(.
Interspecific mrating experimnts- c a:Co.g the three species of
"7ui e-ius were conducted in 55-mm=r polystyrene Petri dishes containing
0.75 per cent water agar at a depth of 4 mm?. Nine plates containing
ten males each (three plates per species) were prepared and supplied
w~ith specimens of ?-n rella61'S "adivivu1s to serve as prey. Three plates
contaicing ten larval Zemales each (one plate per specles) were prepared
anld led as above. After the females in each plate molted to the adult
sta e, t,:ey were trnfre i ruso three to a plate containing
the males of their own specils, the males cf the second Lpeies, or
the males of ,he th;i:6 species. The plates were alternately observed
for 12 brs, after which the females were transferred to individual
plates aid observed periodicallyT for 8 days to ;etermine if any eggs
r...er CrOduced. Subsecuent to the removal of the females to individual
plates, three virgin females of the same species as the males were
place in each plate with the r:.ales to assure thu.t thne males In, eacrh
dish were capable of reproduction.
;eeding. was observed In s-tckc coloniles thiroughout the above studies
by; in-troducing various ;ray neaoe;;Lrs w;ho spc~limens of acco .
three species of Bor.l-1e-is undeJr conditions~ provided 'oy thei ,joe
techniques. Data on competition cf" each s;pecies of 3- t i ls wirth
iiMononchojides sp. and Mr'yle lu-;s s.1tti~': were recorded~c 3y i:-oculating~
plates of agar with equal numbSers of the two spcies under study at
any one time. All possible combinations of the th' `e 0o
Bu~t] erius with each other and with the other twro general wLere: thus
a ULTS .JO DISCLSSICE
,lerirs- IMcirus n. sp. Plate: I
A population of nematodes, determined to be an undescribed species
of onie genu~s Bujt!-lris G-oodey, 1929, was recovered fromr a wet mixture
of soil, rotting hay a 1 seep manu-e. The surlple was taken July 6,1963
from th~e edge of an? in~undated area in a holding, pen of the Departmelnt
oi Veterinary Science, U~niversity of Florida, Gainesville. The specific
nn~am cisL. lacini= flair wars given to denote the papillate flap
cove;-,g the cloac3al opening' of -the male.
The caudal filamenrt Is frequen~tly bitte;- or b-roken off', thus each
of ,he lengths were measured from the lips to the base of the filamen~t.
Femle:(Nl = 10) L, = 1.6 an (1.3 1.8); a = 32.1 (27.5 36.0);
b = 5.17 (%.3 6.0); c = 15.58 (11.0 17.75); Vr = 17.954.51
al:(N\ = 10) L = 1. ,6 mm, (1.35 1. 7); a = 3Lc.7 (30.0 ?42.1)
b = 5.22 (4.90 5.60); c = 14.06 (:3.5 14.75); T = 59.2fo
Body- taperin~g slightly an~te;iorly and conside`rably posteriol-ly,
toe tai il oft~olh sex~es with- 10ag caudal lilam.et. Ciut cle: with fine
,ransverce striatin lateral field indistinct but with~ th-ree lirYs
Subcuticle marked wlth transverse rows of co-spicuous punctations in
,'he head and rack; region, ch~a.ging~ to a rooette arrang~er ment in the body
region, back to transierse rows postanally and ending abruptly at the
base~ of" the cauda~l filamr.~nt. Amnphid opening a tra;.sverse =val located
at the level of the apex of the dorsal tooth. Lip region not offset,
flattened ante-iorly buc. i-,hn 6 membranous lips partially irnerflaced
with the flex~ible chellorbabdians arching up and inward to form a cone
around the oral aperture. Six setose cephralic papillae surrounding, the
lip region, ma~le wri-,h doubled pjapillae in subdorsal and subcventral
positions for total1 of 10. Mvale stomra 3 times as long as w!ide, female
stoma slightly~ broader. Chiotmcm sing anterior 1/3 of as~terior
chamber, cho-ilo=:habsion~s fusing. wJith prorha'jdions at the ;evel of the
bases of the payilla. PrornLbalons encircle the sterior 2/3 of the
anterior chamnber, base of prornabdions attached to mesorhabdions by a
hyaline, flexblSe ring of variable le gth. Ease of nesorhabdicrs rest-
ing on broad metarhabdions which dorsally bear a large cupped tooth,
which is perforated posspically by dorjal gland orifice. -Oth
jsbven~tral metar~habdlon~s bearing a srraller ccapped tooth. Telorbabio ~c..s
fomng basal pa of tee-" `r higly muscular oescphag~eal colla-r,
enlarged dorsally, surroundiing stonia cto the base of .nesorbabdians.
Procorpus swollen anteriorly, then becoming cylindrical and shelling
again to form the valvulated me~zcorsus. Entire corpus strongly
n~uscular. Isthatls broad, enlarges gradually to form~ the broad
terminus wh'rich is no, distinctly bulocus. Tissue of posterior oesopha-
gus largely Eglandularr but intsrspersed with weakc musculature~. Ner.e
rin,- zcrses is An~s jus-t po,t :ic nt m,accrpoo. Ylexcetar pore
vent~ral at level o, r --;e ring. Phasmids plr~leminet i; bcthr sextes.
Fe-al~e. G-onads paired, opposed and reflexedj to i-,ar ;he level
of' th-e vc;gina. Cvaries .sually equal in len-gth, only one egg at any
given time in each of the uteri.
Mlal~e. Testis single, reflex~ed 1-1.5 body diamreters. Rudimentary
bursa discernible on some specime-s. '2 pairs of variably positioned
caudal papillae (3 pair preanal, 1 pair o: very s,-all papi~lle on
cuticular flap covering cloacal opening and 1 pair on posterior lip
of cloacal opening or slightly lateral to that position). Remaining
papillae in typical diplogasteroid arrangement. Spicules paired,
ventrally arculate, capitulanL~ cephalated, calomus narrow near capitulum
tnen broadening and narrow~ing to the lorng ;amina which mnales up nearly
1/2 the total ,enth of the spicules. Gube;rnacuLum 3/4 as long as
5-p-icles and 7 times as long as deep. Dorsal end bearing a scoop-like
p-ojecticn, separated fro ;he remainder of h 'enclmb
septum: and dircted loward the u-de=side of spicules. Ventral 1,'3 oi
gubermaculum enfoling _picule ,anina and bearing two pairs of
The females of Euit-e ins ecianss are distinguished 'oy cat aired
;acven-tral teeth and perforated dorsal tooth which it has in c~~-ommo
c;;ly with ., f1licviev .- T;;e cephalic papillae of B. laei us~ lacki
the pair-ed b;-is"es borne on cthe cephalic papllae of 2. f~ilica-udatus.
to I.. LI- -
^u.r -l em lt:1 D c cet
alerallaerl. G. Feal,
H. Fml hayo r
Males of E. 120-5-'3 are disti;,guished byr the papillate cuticular flop
whil~ch- covers the cloacal cpenilr,i by the gubernaculVumCI whichV bears a
ventrally pointed scoop on the dorsal and and 2 pairs of" cooks on the
venlral end, and by the 12 cadal paplllae.
Single females on slide no. I labeled "Euit'rler'19 !Da;nin (holetype),
in collection of' the Departmenit of Enatomclogy, Unriversityy of Florida,
;=eposited in collection of the Department of Entom~ologyr, Unilversity
of Florida and others In the unitedd States Department of' Agriculture
Nematode Collection, Nematology Investigations, Agriculural Research
Service, Beltsville, Marylard.
Game a. le-in lac .s 3.-as recovered most frequently from
semiaquastic habita.s an~d appeared to be we~l datedd to this typSe of
environment. Ur~like most soil nematodes which sirk rapidly in water,
J. 1 cin ai was repeatedly observed in water of a depth of 2 to 3 cm to
swim from the bottom to the surface and remain rear the surface,
swiminglr, strongly for periods in excess of 15 min. Other individuals
were observed~ swimming just beneath the suirface of the. hatED in a
^0 gal ;;aqua~riu 3 1Lays after introductions,. Th~e sp~ecirens were then
e~aer, cy t;,e fion. 3,10:;5~ Tes woe main-,ained fo; a year i,; Petri dishes
fi_;ec w,,;ith ater, jfter wh~ich time the technrique wars abandoned in
"avor- of a starndardizec agar m~edium~n. Swimm~ning wIas accomrplianed wcit'
a whipping motion of the~ entire bo~dy. Swimming, was frequer,,_.y in-te-r-
rupted by a pause of 1 to 2 sec. During these pauses the head, with
lips extended similar to the attitude illustrated by Co'ob (2) for
no-chus, was mnoved f~-rom side to side in a je;-kin-g or groping motion.
Pauses in swjimrming, became longer and more frequent w;hen to~ specimens
iwerle striuilated by intr,-oduction. of --7na-rzllusu redivrivus or other prey
nematodes. On-ce swimm~ning activi-,ies were resumed, the ilps usually
resume-,d a closeC conical position. While swimming, E. lacirus exhnibited
an apparent greater degree of control of both muscular activity and
direction than is see:: in t:,e rather frantic flailing of the body by
slch aquatiz nema~todes as species of Pris-.-toi'la ms.
Numerous samp~les containing specimens of B. lac~nus were obtained
fr'om Mr. R. P. Esserg *31ivisn of Plant Industry, Fcrida State Depart-
m~en~t of AgricultLre, G-ainesville. Th~e samples w~ere collected: from a
variety of locations, including nursery soil and comme.rcial earthwormr
beds. They provided lor frequent comparisons of wild populations and
use stocx coxonles.
Emb-v ---. total of 19 eggs of A. 'acirus wras used in two
different embryclogical studies. ;he eggs ranged from 68 to 81 p i
length and 37 to 43 : in wJid-h. average time lapise from ovlposition
(ranges included pare~nthtically) to each developmental stage appears
in sema~ry form In the legend of Fig. 1.
Eggs were deposited singly while in the one-cell stage. Eggs in
the two-cell or later jtages were obesrved 1n th-e u-,er- of actil,S
fem~ales and those that had been, injured or ha~d expe~rlizzed a mechanical~
blockage of the vagina o;- vulva. Su7h far. 163 die prior to advanced
development of thu;~ e-g ,.ne p'-7 - waZ. ;v` ve o;`'El b Jrd -o
this species. On twrYo occjlailns, ve;ry oldj -t.:ales wer~e casero-~C, playing
eggs without a chnorion (3ig. 15).
Cellular division pr-oceeded rapid -, 10i teo. -n=ra;
pattern described for ebr'oyonic dev;elopmrenrt of mother ne.:atdes.
notable aspect of the intrachorionic developments of -. 1-ines was thie
low level of develop ment of the sto,al rhabdlins the first ste
larva at the initiation of t;-.-: first n~o~t (FIg. ;Im). At th-,s devel~o,-
mental stage the rhabdiorns were so pcoorly developedi as to be ir-defir.ole
within the chorion and wore resolved w~ith difficulty even or larvje
whic had been~ removed fromr thre chorionr. Thnus It was necessary -to me~
the separation of the ,a'oial cutile (7i 1ls), actually ,rn irnic tijr
of advanced stages of mroltling, in; 'ate- larval stages, as ir.diatlive of
the initiation of the intractorionic molt.
Larval egress fcrom the e,6 :as a=cco p~lsh-ed ,y mos, 1:vaca ar
ently solely thrug reae st`echin oftz corca Al bsn
l ar v a e re p e a t e l p r e s t he i r l i e i s g i s t e c c i a ; t i
resulted In the chorion, becoming extremely flexiole ~jus pric: ,o e re;s.
At this point, twio of' the larvae were observed3 to idrra -:; c:;;-crio IP;int
their stomna while vorsy vn te dorsal tooth; :.0 cozzact,
could be observed, however;-, betwJeen. te tcoo~-Lth CandO choiae -
to 60-h-r period from? the first la,-al molo -uo agre~s~ prjved~ To 3be ,:-
mos variable period in1 the develop mentci of the~ egg of. 1 M (?-eig
ip.The m~olted cutileo the first stager 1&:Ja could 1;ot Le co-Seved~
in any empty cho;ions.
Fig 1.Elmbryonic development of Butlerius lacinus.
a. Egg deposited without chorion. bo. 1-cell stage
intrediately following depositlion. c. Initiation
of' cleavage. d. 2-cell stage (.5 1 hr).
e. 4-cell stage (1 2 hr). f. 8-cell stage
(2 5 hr). g. 16-cell stage (6 7 hr).
h. 32-cell stage (7 8 hr). i. Morula
(8 -10 h). Blastula (11 15 hr;.
k. Gastrala (14 16 hr). 1. Tadpole (15 17 br),
ncte poorly developed rhabdians. n. Molt (25 -
30 br), note loose labial cuticle. 0. 2nd larval
stage (30 38 hr), note rhabdian development.
p. Eclosion ;38 60 br).
Tale 1. Larval Stage Duration and EodyT Langths
of Btil;e-ius 'aci-r-s
Bodyr Length ini mm
S we e Du st~ion 1 ? s nvear Ro'
1st 19 0.30 (0.30 0.31)
2nd 39 0.50 (0.35 0.63)
3rd 17 0.76 (0.68 0.83)
Oth 21 1.05 (1.oc 1.07)
S 01.3(0.95 1.53)
FollowingX oviposition,, the adult stage wras reached In an average
of 114e `rs oy r'emrales and; 102 brs by males. Longevity studies pe-r se
were not conducted, but maximum~ adult longevity was detern-ined to be
in exccess of 6 wk~;s for virgin irndividua~lj. The reco;-ded time. schedules
1 e 70 .The duration of' thei various larval jidages of
B;. laci~n-ls was, for the purposes of this study, cons,~idered to oe the
timle lapse from: the initiat~ion of one mrolt to the next initiation of
mcitingc. ;aral; len~ths were late, calculated using specimerns frorn
stock colaries, the stages o' development of which were determined on
the basis of degree of genital primordia development. Sex of the
larv-ae can be determined with certainty in the fourth stage by the
elongate posteriorly directed enlargement ol the male primoraium as
compared with t's balanced, more robust primordium of the female. The
larval stage durations, reported in Table 1, are averages of" 19 indi-
vJiduals for the first and second stage (to egr;,s f'romn egg) la;vae and
17 each for- the male ane female larvae of the post-egress second,
third and fourth stages. Lengths given are averages of ~5 individuals
with ranges ircluded parenthetically.
profibly ajproach th,- m~iniumr since -,he duration of eace la~va- stage
is depende-t to a large legree trpon tse availability and quality of
food. The body length measurements, however, probably are similar to
specimens produced in nature, since tce larvae that wrere measured had
developed under the competitive environmental of a heavily populated
:03ir -. Prelimlinaryr stuc-es of the molting process in B. lacin~us
indicated that tne sequence of rhabdion migration and formation is
identical to that of B, pa~sc--us, given later.
P arrod-ction. Males of 3. 12cirus were found capable or' locating~
virgin females at, distances u~p to 80 mm- wJhile in 0.75 per cent water
agar plates. Immrediately following their introauction to a plate
containing virgin females, the males appeared to uncergo a period of
disorientation, rain fro a few seconds to, in a single instance,
10 min. During this period they moved aimlessly about, moving their
cephal?~ic region fr~om side to side in a rapild, jerking ~c~ion. This
motion was originally thought to be a response to the proximity o;
prey; howrever, it became quite apparent during~ the rep=oductive studies
that ,his behavior is a positive tactic response to any attractive
stinuluo,. Th;is behavior pattern is herein referred to as castin~g."
Thre period of disorientatiosn wJas termi~nated when T~he male moved
-apidly in the direction of tnz female with the casting being some-
what subdued during periods of rapid forward motion. The female was
approached in an approximately straight line, even in instances where
masses of prey nem:ratodes ~e-e: be~rreen the male and female. Such
cbs-;ctl~rucis rarely resculted in any apprteciable delay or deviation
f~ronz the line of apjproacs. -Th at-tractan~t appeared to origi,.at-
the: vulvar region, since iniStial contact was made there if the femarle
did noti move d;;ring the apprach: of t;he male. In, irstance~s where the
female had vacazed a location immer.diately prior to his arrival, the
male would initiate a very rapid casting and then follow the path of
departure of" the female. Und conditions of continual movement by the
female following the introduction of the male, the li-e of approach
formed a curve to intersect the path cf the female, and ultimately,
the male followed the female until contact was made.
Ipon contact with the female, the anterior part of the male moved
beyond the vulva until the caudal region of' the male contacted the body
of the female. The caudal region curled rapidly around the body of the
female at the first Doint of contact. Considerable variation occurred
f~ollowLing the '.nitial approach and contact. Most males initiated a
thnristingJ of the spicules while coili-g and un~coiling around the female's
jody. Other males did no, thrust therr spicules but lay loosely coiled
,ntil movements by the female brought her vulva into close proximity to
the male's cloacal opening, at which time the male would tighten the
coil usually with sufficient force to constrict the body of" the fermale
When alig-nment of the vcalva ane cloacal opening was achieved., the
longr spicules wer-e Inserted rapidly, frequently to a depth exeed~i-c g
that of the T1:gina (Fig. 2b). Th~e a 16 of the gubernaculum was also
inserted into the vagina (Fig. 2c). Th urtono the union was
exceedingly variable, ranging fron 2 to 30 min, during wh~rich time the
vagina and uteri underwent a muscular activity hihrsutd na
foservaole mov-ement of isperm into both ute;-i. Th~is muscular action was,
in most cases, sufficiently forceful to bend the thin laminae of the
spicules (Fig. 2c).
Subsequent to copulation, the female moved awray, leaving the male
loo~sely coiled. Frequently, if the female moved awray before the
spicules and gubernaculum were withdrawn or if the female waas disturbed
during copulation, the male was dragged by the female. Apparently,
this resulted in the inability to disengage the sclercnized gubernacular
ho~oks f'rom the vagina. Fe, ales in liquid media frequently w~ere observed
swimming vigorously with one or more males coiled tightly around their
Cach~ virgin femrale of B. lacinus copulated an~d p~rodul-cd viable
eggs whten introduced to plates cca;taining males of' this species. Nio
at~temrpt was made by the ma.les of' the other two Eutleriu~s spp. to cop;-
ulate wjithn females of B. lacirnus, nor were the B. laci.us males observed
to attempts copulation with females of the two moncelphic species.
Upon contact with them, they eith-er withdrew rapidly or killed and are
tm.The females of the moncde-lphic species deposited sc eggs w~ithin,
8 days following separation from the males of B. lacinus.
F~ edi-e. Specimaens of' B. lacinus usually feed by ingesting small
prey r.ematodes in toto and by breaking the curlice of" larger prey
nematiodes and ingestinig the body contents. Small nematodes, such as
Cyrli-nfracorous and '3robel-inians whten caught in the midsection, were
con~sumed by b-eaking the body wall and then ingesting the erntire body.
Ingestion cf whole prey specimens Is accomplished by retracting
tE, do~rsal tLocth (and presumably the subventrals), dilatir.g the tri
cdiate lum~en of the oesop ;agus and moving the entire prey body; through
F~ib. 2. Copulation? of Butlerius lacinuls.
a. Copulatory posture, note constric~ion of
female' s body in vulvar region. b. Spicule
Fenetration. c. Bending of spicules due ;o
femrale .luscuilar acticn, nc-e depth to which
gubernaculum is inserted.
the oesophagu~s by a perijtalsis-ilik ucua cio.Drk;t
activity, secretions moved in the dorsa;l oesophtagal ;land duct,
located dorsally in th~e --ad-iating musculatu-re of the oesoph=-gus, and
into the posterior por,ion of the do=-eal tooth1 (Fig. 3:).Se,;io
of the dorsal oes3phagesl enzymrres thr~ough:- the~ -oo :; c-r ice w.as
observed during fecnbt secrztio,?s dId mrov, throuJ. L,, thiscrifce
accompa~nying dorsal tooth ::ov~emen in compress slides. Sclerxled~
structures, such as the rha'odions, of lar-vae uf ? 2 : a--~s were fro-
;uently observed in the intestines of adl 6eies Fg 3b).
Thie body wall of prey su~ch as ? E ~-elle_1s .-=ji.' i; r=;l;tjaFe
by E. lacin~us 'oy the use of the doroal toot,;. Temmrcslp
pressed against th-e 'oody wall of the pryan husoa elo;-gaTtes
convulsively pu~lling, thecutie (sull b mu~ltple pumpings) to the
level o,' the dorsal too,:h.
Under ir. vita cc~idition~s th;e prey of' _.1,.s nl-,
Acrobeloides sp., Zreviibncea~C sp., ~-'ti~ i.._ re osp, ?-rol
-redivi u:s, ParaTrol~ irus sp., and Rhr'ri--i sp.Te niene
cannibalism in B. i~lrcirs was low in welli-fed colonies, bit i, unde-f;d
or overpopulat~ed colonies cannib~alism; ozccurd fr'equently..
Survival ability of r. 1Ircnus in compejti~tionr wri b,-. jtheotr1.
and water colonies was determrinied by colonizing }. 1:'--us writh a~ch cf
the others. B.lcaproaba'ol dUs ,o Its g rea-ter deg;ree cI aciv
and larger size, destroyed the populations of the other two z,; leries
spp. in 'ooth water and agar colonies. In water colonTies, probably beca;se
of its abilty, to swiJ:;, B. Tsr niaiepoltIo t blj wrhen
coonze ithete l:hie po 1. -3 i, a ti
Fig. 3. Feeding of F:;lerius lacinus.
a. orticr, c, ,,ne intestine of an. adult
containin, part of the stoma and oesophagus
of a larva. b. Adult head of 'emale, note
dorsal gland orifice and articulated
observed to prey on the larvae of eachr. Under conditions o,' agar
colonization, both De-vrlasmus subtili~s and Mnornchoides sp. destroyed
the populatviors of B. -arin 5
Eutler`iusj na .splus n. sp. Plate II
A population of enemtodes, determined to be an undescribed mono-
delphic species of the genus Butlerius G-oodey, 1929, was recovered from
a sample of moist frass of the horned passalus beetle (Perilius
fisuncus(Illiger)) in an oak (Ouere j sp.) log. T;-e sample was
obtained June 11, 1963 from "College Park," University of Florida
campus. The specified namne r ;s-,lts is given to the nem;atode to denote
t;he type locality. The caudal filam~ent is frequently missing, thus all
measurements were cased upo;- lengtn from the lips to the base of the
Female: (Nr = 10) L = 1.08 mmn (1.0 1.25); a = 22.04i (22.0 22.7);
o = 4.28i (4.0 4.5); c = 10.8 (10.0 11.25); V = 23.0568.3%5 (66.6 -
Male: (N = 10) L = .81 mmL? (.70 .95); a = 19.5 (18-35 22.50);
c = E.4~0 (7.0 9.0); T = 48.614$.
Biody tapering a:.teriorly and posteriorly, greatest body diam-eter
at 0.1ry flexure, both sexes bear an unusually long caudal filament.
Cumiole of bot:, series bearing longitudical ridges broken 'cy fine
transverse st-iae resulting in a weakly beaded appearance. Subcuticle
exhibiting punctations between striations In transverse rows, puncta-
tions occupy alternate positions longitudinally, a character observed
with difficulty in the males where the beaded longitudinal ridges are
more pronounced. Amhid opening transversely oval, located slightly
anterior to the apex of the dorsal tooth. Head not offset, somewhat
blunti anteriorly wiith corona of 12 recurved, membranous lips. Lips
innerfaced by cheilorhabdions to point of recurvature. Cheilorhobdions
,rticulated on anterior prorhabdions. Prorha'cdions joined, by a flex-
Ible membran~e of variable length, to the outside of the :,esorbcabdions.
?Isorhabdions broradly based on the metarhabdions. A large tooth is
present on the dorsal mnetarh~abdion and a cup-shaped rasping plate is
present on each subventral metarhabdion. Dorjal tooth perforated sub-
apically by dorsal gland orifice. Rasping plates each bear a single
semicircular row of 12 or more denticles. Female stoma twice as deep
as wide, male stoma 3 times as deep as wide. Female with 6 heavy
caphalic setae, male with 4 additional setae located in subdorsal and
suoventral positions. A swollen oesophageal collar surrounding stoma
a; far as base of the mesorhabdions. Procorpus narrows poste;-ior to
base of stoma. Metacorpus swelling to form valvula",e median bulb.
Entire ccrpal region strongly muscular. Isthmus rather broad, gen~tly
expanding poster-iorly without forming a distinct basal bulb. Isthm~us
and posterior region of oesophagus predominantly glandular but inter-
spersed with weak musculature. Nerve ring crosses isthmus just
pose~rior to the ;netacorpus. Excretoryr pore ventral, equidistarnt
oetwreen 'case of the metacorpus and cardia.
;TAnale, M~on~odelphic reflexed, lininrn of vaeir~a uYe_rin
peculiarly refractive. Post-uterine branch usually filled with sperm.
Ml.Stoma laterally depressed wrhen viewec en face. Tests
single, anterior end reflexed from 1 ru 2 body diameters. Nonbursate.
pairs of caudal p~apillae in typical diplogasteroid arrar.~ngemet,
variable in position, 2 to 3 pairs preanal. Spicules heavy, paired,
ventrally arcuate. Capitulum; cephalated, calomus broad and narrowing
to the- long lamin;ae which? corrprise more than half the total length of
the spicules. Tips of laminae gently arcuate enfolded 25 per cent of
t.?eir length by the gubern~aculum. G-ubternaculur.~ keel-like, 40 per cent
tne length of spicules and half as deep as long; tip and subventral
margins heavilyr sclerotized; dorsal ma gin collapsed and very lightly
sclerotized. Bottom of gubernaculum~ bearing large lobe of relatively
FCemalzs of' Bu~.~rleins raCSsfus are distinrgCuishes f-rom all didelphicl
species by the single ovary. The single large dorsal tot,;; whrich Is
perforated subapically by the dorsal oesoph~ageal glan~d ori ice and the
rassing plates located on, each subvent;-al metarhabdian separates P.
Dessal~s from B. mrnbysters for which Taylor (16) illustrates one large
,nd one small dorsal tooth and a single, semicircular rasping plate.
!;ales of B. rcassa~lus are distinguished from all other males of the
genus Emlariu-s on the basis of the u~nicue, keel-like guberncou~lum.n
.~~3 Mae aetal iorl
Single ;' ;Le on, s~lde: no. 1I labele~d "Eu'lerita,:Ll3
holotypee), in collection of the Department of Entomology, University
of Florida, Gainesville, Florida.
Deposited in collection of the Departmenr t of E4~ntomol; og, Ui'lversity
of Florida and in the United States Department of Agric iture :;ema.todec
Collection, Nematology Investigations, Agricultural Researc erie
frass of the horned passalid, in th yelcaiyadto te oa
tions in Alachua Cou~t~y, Florida. So spcmn eercvr =0
numerous other organic substrates; a definite rabitat spjelcifctyr,
therefore, must be attributed to this sp~cies, ev;er though, ,elatlively
few~ samples of passalid f'rass wer-e processed. u
although capable of swimming, lacks ,he obvious ;adapta in to acuatic
conditions noted for 3. lacinus~. Altho, gh spucimen,~s o bt pce
employ the same basic m~ovem7ent in swjT;?inin, Z. --re -- swims~ wiLt: a
mnuch slower and more exaggerated rhythmic moemn of thr. body a:-e does
not swim above the bottom for extended periods of time, as does 2.
in -ength~ and 37 to Lc3 ; in id:h The-CU choic o th~e cagga x%;ioieG
hyaline, adhesive protube;-ances (Fi;. Ik and s), the; functionr of hirlch
was not determined. Average~ time lapnse (rangaes included parent~etica-ly/
from oviposition to eacn developmen~:tal stade Is summarize-,d in tie
legend of Figs. 4 and 5.
Eggs were deposited singly while In ,he ons-ell say j2,~ _o
the two-cell stago, .;ose in more advanced s-tages of 6,~velay:.u:.1,-~ C
were not observed in the uteri of healthyI fem~ales. Egg deposizio- wa~s
not observed for females bearing more than one egg or an egg developed
beyond the two-cell stage in the uterus. Such 'emnales often lIved for
considerable periods of time, however, and E' i7tntakis I-riC-da, wrs
Th-e em'oryonic development from ovipositio- to eclosion 0: '
?assnlus required nearly twice the tinme dete;mined for -. ?
Egg development in B. ?anala:; followed~ tC- saegeealpo -u thTe
embryonic development described for othe:-; rnemoatods sno 2:E
lacinus, the stomata of first-stage larvJae o' I. r--` ce utibited a
very limited degree of rhabdional formatio Th naco`i
molbting process was more easily observed in r. rn~ : beaue o~, f
relatively greater degree of inflation of the seaatn cephali
cuticle (Figs. Sh, i, and p).
Larval egress from the egg wras apparently acc=enplsishe 1, t;.,
same manner as described for B. lacinn"s, buu, the 70 to 73 br variation~
in time from the first larval molt of 3. -.- e in~ to cegress ;as ; Ss
thian that reported for g. lot'~..ns
Newly emerged larvae were observed in ,;wo case3 to re-2nzer the
Fig. 5. Emioryonic development of" Butleries ivassalus f'romn
blastula to eclosion.
a. Blastu~la (19 2+ hr). b., c. Gastrulla
(22 28 hr). d., e. Tadpole stage (28 -
31.55 0.) f. 1st larval stage (32 4-0 i;,).
g-1. Mlti;.g (38 49 hr), no~te separation of
cephalic cuticle and degree of rhabdion develop-
mnent. j. 2nd ;arval ;tage (47 60 hr).
k.2nd larval st_=e, note ventral view of stoma
exh1ibiing dorsal tooth and subver,,ral rasping
plates. 1. 2nd larval stage pressing against
chorion. m., n. Egress (70 78 hr).
o. Charion after egress, note molted cuticle In
lowerr right corner. p. Mloltin~g larva which cias
been removed fromr the e g, note sepa;-ated
cephalic cuticle with dot-like mol~ea rhabdions.
Tabl 2.Larval Stag- Duratiion and Bo~dy LengtLhs
of Butlerius p asalus
Body Length in mm,7
Stare Dur~a tion in hnrs Ave ra ge Ranfge
1s, 11 0.30 (0.28 0.32)
2nd 66 0.14 (0.38 0.58)
3rd 45 ;.50 (0.50 0.60)
LAth 6, -9 0.68 i0.60 0.72)
53 0.70 (0.60 0.80)
The adult stage was reached by females in an average of 212 hrs
and by males in an average of 203 hrs.
01;oti:T. Th;re molting process was initiated w~ith; a quiescent
period of variable dur::;tion. During this time the wJalls of the 1lamen
of the oesophagus cleared and the procorpus appeared to withdraw pulling
withl it thie posterior chdnmbers of" the stom~a (Fig. 7a). This resulted
in t;,e separation of the prort~abdions from? the mesortabdions and the
stretchirg of the elastic connection membrane (Fig. 6a). Following
sep~araion of the pro- and mesorh~abdions, a gentle side to side move-
ment of the posterior stoma was initiated and the cephnalic tissues were
slo:;ly with~drawnm (Figs. 6b-d). Slmultaneously, the tissues surrounding
chorion ans maul the molted first stage larval cutlile, whico :emained
in tme egg follow~ing egress (Fig. 50).
Life cyrc~e. The duration of a larval stage was considered to be
the time from initiation of one molt to initiation of the next. Larval
mreasurements, obtained in the man-ner described for Ba. lacin~us are
included with larval stage durations in Table 2.
th-e posterior stoma became hyalin~e (?ig. 6c). Thie posce-ic-r stars-.
ceased movement and contracted slightly ,s aint outline; of the newr
rhabdions formed outside and slight;y posterior ,o the old c.-es. A
this time the rew lip reiG apeae 8 (;=-,). AS the 8 att28~ Of
the stom~a became easily; discern;ible and Y;he chatilo- a-d pj-:orkbu,ns
formed, the elements of the larval stor;a; and c: ohga lizia~~bC~- 11_;,
migrated almost imperceptibly anteriorly and the lining of the
esophageal lumen regained its normal appearance (Figs. 66-,). .-ky---
ment was resumned upon th<~ -ilerotizai -; ol the new ;n~i~;rhaud an, C.:Zio ,wre
continually flexed for the remainder of ecjysis. "he body cuticle
began to stretch a;, loosen ~i~th' increa;3d motion by t~F he Tlo A
the mass of larval rhabdions and oesoph~ageal ,rlnig ;ire 5_cutie -.::.5
the newly formed stom~a,-the linings of t,,e amphiidial pwc~e-;s wo-ri-
withdrawn (Figs. 6h-j). At this stage, the lipe of th~e vul~vr, were
discerniole (Fig. 6k). hs -the n~ema.tod: pushE~d and mauled the jce
rlhabdions (Figs. 6j and 1; Fig ?') teLnig o h exic^etar; ,ore
(Fig. 6m;) and rec~tum~, wrhich were the onl~y attachmnent points fe:- LL
loose larval cuticle, w~ere molted by rolling the bod wilthi:; tl; l,--ial
cuticle (Fig. 6n). Once all attachm~ent to -he larval cuticle was
broken, the young adult repeatedly pre:ssed against It in -theL cevicaLr
region (Fig. 60) until ecdyisis wVas compr.leted 21; to 35 5:3 are: the
first signs of molting (Fig. 6p~; Fig. 7c). Newly molded Ic,iviou:18
were occasionally seen to return and maul the moltec curicia.
Reproduction. Males of" E. TIss-;-1s located virgin femralus i.
0.75 per cent water agar p lats at distnc~i es up to 80m. ecr.
appeared to undergo a period of aisorienttio 'L o inrdcis.t
Fig. 6. Mlolt~ng of Lutler'.us D~ssal s.
a. Initiation of molt, note separation, of pro-
and mesorzabdions. b. Withdraw~al of cephalic
tissues. c. Eyaline rhabdion formation area
developing in p-rocorpus. d-g. Dorsal rhabdion
fornation. h Collapse of larval stoma. ,j
Mc~lting zamphidial lining and w~ithndraw~al of' larval
stcma. Vulvar development. 1-n. Mauling
larval rhabdions. m. Flexing of" larval cu-ricle,
note larvali anus located dorsally. n. Mclt of
ex:cretory pore lining. o. Pressing against
stretched cuticle. p.Cmleino ecdysis,
note complete larval stomna fc~right.
a b c d
e f g h
i k~ 1 I,
Fig-. 7. Molting of Lutlerius cassalus.
a. Initlition of molt. 'o. Pressing against
larval cuticle prior to egress, nore rectal
lining appressed to cuticle. c. Comp-letion
o csinote larval cuticle and molted
plate containing a virgin fema-e. Followlng the period of disorientation,
the a n6es initiated ii casting be savior which was much more subdued than
that observed for males of B. Incinus. The approach, though less rapid
than that of' B. la-;..n 3, was equally dir3cLL :t, and apparently no more
difficulty was encountered 'oy obstacles such as masses of prey nema-iodes.
Observations indicated a conditioning of mated males, which resulted
in their achieving union with the female more rapidly than non-mated
males. ipon contact with? the female, mated males m1ost frequently pro-
ceeded beyond the valva and assaued a copulatory coil only after the
caudal region made contact with the vulvar region (Fig. Sa). The non;-
matedmales howeer, m re f lreqenly assumed a copulatory coil immedi-
ately up-n encountering the vulvar region, resulting in initial cop-
ulatory attempts in the cervical or anal region of the female (Fig. 8b).
'The gubernaculumr was not observed to enter the vulva, and the
laminae of the spicules were not observed to penetrate more than 1/2 the
depth of the vagina (Fig. Sc). Following copulation, a gelatinous
appearig plug was observed covering the vulva (Fig. Sd:. The Function
of the plug was not determ-rined, but it could function as an adhesive
during copulation or as a sealant to prevent ser~m loss immediately
following copulation. The plug was usually lost within 5 min after
The interspecific mating tests yielded no copulatory contact of
males and femle ofB sa'u~s with the other tw..o species of Butle~rius.
Conversely, all intraspecific contacts resulted in production of eggs.
~eedi- Specimens of B. passalus fed readily upon the same prey
reported for 3. lacinus. The cuticle of large prey nenatodes was drawn
Fi 8. Copulation of' ?'.tleries me **i9LUS.
a.Initial coulation attempt by previously mated
Cales, note prox~imity to vulva. b. Initial
copulation attempt by non-mated males. c. Spicule
penetration, note position of" gubernaculum.
d. Vulvar plug.
F~ib. 9. Feedinrg of _u leriuss russalus.
a. Larva attackiing Panwcrellus redi~viv;us
larva, note rupture In cuticle resulting from
previous attack by same larva. b. As a,
no~e, P. I~j'rdivius cuticle drawn to point of
dorsal tooth. c. Ingestion~ of prey 1in toto,
note dorsal tooth wiithdrawn into wall of
i;,to the stomai and ruptured w;ithn the dorsal tooth (Figs. 9a and b).
S;-aller prey ne,,Ltodes were ingested In '7te by fodldng the dorsal
tooth into the wall of the stoma (Fig. 9c).
Completition studies indlicted that ~. nassc lu is the least
competitive of the three species of Butleri~us under study. Bultle-rius
majsalus failed to become established. in mixea colonies with B. 12cinus,
FMznonrhcides sp. or Dorvlairus sub lis. In mixed colonies with the
other monodelphic Blt~lerius sp. they became established, but, after a
period of inadequate food supply, they were dominated by that species.
Euuleriuis hame~spicts n. sp. Plate III
;i popculation of n~ematodes, determined to be an undescribed mono-
delphic species of the genus Bu-lerius Goodey, 1929, was recovered fron
a sample of wet organic soil near the overflow drain of a spring house.
Th;e sample was taken July 13, 1963 from the property of' Mr. J. C. Riussell,
Scnford, Florida. Th~e specific n~ame h;rosliculs L. hamo~ = hook -
L. sic= spicule is given to denote the ventrally arcuate spicules of
the rmale. All measurements are based on length fr,;m lips -to thre base
of the caudal filamient since the filament is frequently bitten or
"emale: (:: = 10) L =.93 mm (.85 97); a = 186 (17.0- 95);
b =4.4(43 -4.); =11.3 (9.7 13.0); V = '67.2(a (64.7 68. ).
El: ( = 10) L = .77 mmr (.67 .85); a = 31.1 (23.9 38.8);
o = .,2 (3.9 1 .8); c = 8.7 (7.8 9.7/; T = 39.7fe.
Body tapering anteriorly and posteriorly, greatest body diameter
at ovary flexure, bo~h sexes bear an unusually long caudal filament.
Cuticle of both sexes bearing longitudinal ridges broken by fine
transverse striae resulting in a weakly beaded appearance. Subcuticle
exhibiting punctations which are evenly spaced in either longitudinal
or t;ransverse rows. Amphid opening large, transversely oval, (larger
in males where they occupy up to 1/3 of the diameter of the head),
located anterior to the apex of the dorsal tooth in females and at
the level of the dorsal tooth in males. Head not offset, bluntly
rou-ded anteriorly with six men'oranous lips which are partially inner-
faced by cheilorhabdions. Cheilorba'odions broadly articulated and
overlapping the anterior prorhabdlons which are enlarged near the point
of articulation. Prorh bdions joined, by a flexible meambrane of variable
lengh, o the outside of the mesortabdions. M~esorh bdions also heavy
and broadly ba ed on the metarkabdions. A large tooth? is present on
the dorsal m~etarhiabdion and a cup-shaped rasping plate is present on
each subventral metarlkabdion. Dorsal toothr perf~orated subapically by
dorsal gland orifice. Rasping plates each bear an imperfect row of
small denticles reaching only/ half the distance across the plate.
Female stomra 2.5 times as deep as wide, male stoma 4 times as deep as
wide. Female with 6 heavyJ cephalic setae, male with 4 additional setza
located in subdorsal and subver.tral positions. Procorpuis forming a
sli htly enlarged collar surrounding the stoma as far as the base of
t..e metarhabdiuns. Proco-pus n~arrow~s s lightly pcsts-,ior to the stom~a.
Metacorpus enlarging to form a valvulate median bulb. Entire corpus
thick and heavily muscular. Isthmus rather broad, gradually exp~anding
posterorly without formrirg a distinct basal bulb. Isthm7us and
posterior region of oesophagus predominan~tly glan~dular but interspersed
with weak musculature. Nerve ring crossing isthmus just postcerior to
the metacorpus. Excretory pore ventral, just poster-ior to nerve ring.
pose a. Monodelphic reflexed. Lining of thle vac na uterina
peculiarly~ refractive and exhibiting~ a small anteriorly directed puch,
function- unknown. Post-uterine branch usually filled with sperm.
Mal'e. Prostom laterally depressed when viewed en face. TestCis
single, anterior end reflexed only slightly more than one booy diamreter.
Nlon~bursate. 9 pairs of caudal papillae (3 pair preanal) in typical
diplogasteroid arrangement, variable in position. Spicules heavy,
paired, ventrally arcuate, capitulum cephalated, calomus broad and
narrowing to the long heavy laminae which are enfolded 1/3 of their
length by the gubernaculium. Tips of lamlinae strongly arcuate.
Gubernaculumi keel-like, 1/2 the length of the spicules and 1/2 as deep
as long. Gubernaculu~m heavily sclerotized In the seto weei
enf'olds 7;a opicules. The remrainder of th're keel margin. is complete
and equally sclerotized.
? .males of E..' rius haespicu= a;m distcinguished fromr didelp~hic
specis of n-?sr'lls oy the single ovary. Tae single large dorsal
asn ra rr ~ae man, vetr
-^ t1cle T -
tooth whion i3 perforated subapically by the dorsal oesophageal gland
orifice and the rasping plates loc ted on each su'oventral metarhabdion
distinguishes it from; E. manGI-, TE~. Separ,,tion fro: B. passal.uc is by
the half-row of dontcles on each rasping plate, ;..d cy the six lips.
Males of B. hazospcricus are distinguished f~rom all other species by the
ar~cuate lamlinae and the completely sclerotized border on the keel-shaped
Single femnaze on slide no. 1 labeled "Butlenius hamosnicus"
holotypee), in collection of the Departm~ent of Entomology, University
of' Florida, Gainesville, Florida.
Deposited In collection of the Department of Entomology, Un~iversity
of Florida and others In the U-rited States Department of Agriculture,
Nematode Collection, Nematology Investigations, Agricultural Research
Service, Beltsville, M~aryl~and.
corl.B-tleninls harasicus is closer to B. reassics in etholo,3y
and n~orph~ology than to 3. lacint.!s. The movements during swimming as
wiell as the behavorial responses exhi'bited in the reproduction studies
and f'eeding habits are of a relatively sluggish nature when compared
to those of 13. lacinns.
The a7uthor is i:,debtea~ to Mr. R. ?. so o tyn "- ,10,,,
containing natural popu~lations of' -. vos rec7~;~~ovi-er 6 fre the2
same~ types of ha'oita-ts as L'. I ;cir s-. Theje populati .v p~rvid
for continual comparisons, of rild and 1aboratory populations.~
Emryloy.The 20 eggss used in. t;;is stucd:, me~asured fcro 18
87 pi in length aid 31 to 77 p in width. Thle enorio-,, as did ;.at of
B. r.csa:as, exh;ibited a large rnumber of relatively large hyaline
adhesive protuberances. Average time lapse from oviposition (ranges
included parenthetically) to each deve opmental stage is seLr~arlzed ir
the legend of Fig. 10.
Eggs were laid singly while in -,e~ one-cell stage an~d, as reportooa
for B. passalu~s, two-cell or more advanced ;tages of developers., we;re
not o'oserved in the uteri of healthy femlales. Oviposition was3 -ot
observed for females hearingg more than o;,e eg;, or e as developed to
or beyond the tw-o-cell stage, in thel~ir teri; 'out the latter feal~es
did live for considerable periods of tie.
The embryonic developments of h-scapira~ I.as of' on,, 31, sitly
shorter duration than that of 3. pa~ssalus and devrelop.:;3;z wasgrea1
the same. The first stage larvae exhiuited~ thesaelitdraiol
development and the intrachorionic making;; process w~as dir'fliclt to
Larval egress from the egg was accomp~lish~di 3apparetly iin -,h. s
manner as described for the other tw~o sp~eces of Bt3~Leruatog
interval between intrachorionic molting and egresj s as no, so varia'ole
as that reported for B. lacinus.
Fig. 10. Embryonic developed ,t of ;'ut~lerirs harnosiicus.
a. 1-call stage. b. Initiation of cleavage.
c. 2-cell stage (1.5 2.5 br). d.Division
toward Lr-cell stage. e. @-cell stage (2.5
3.5 ibr). f. 8-cell stage (3.5 85 hr).
g. 16-cel; stage (12 15-5 hr). h. 32-cell
stage (1;.5 17.5 hr). i. M'orula (16.5 -
19 r).j. Blastula (19.5 24 hr). x
Gastrala (26.5 30.5 br). 1. Tadpole (29.5 -
35 hr). m. 1st larval stage, note degree of
rhabdian development. n. Molting (44 lc9 br),
note separation of cephalic cuticle and rhaibdian
development. o. 2nd stage larva. p. Egress.
Table 3. Larval Stage Duration, and Body Lengths
of Butlerius narnosncus
Body Length in rmm
St-ice Duration in hrs Ave ,re Rante
1st 10 0.25 (no variations)
2nd 61 0.36 (0.27 0.45)
3rd 35 0.50 (0.43 0.60)
Ith on 40 0i.58 (0.48 0.65)
bc 40 0.66 (0.60 0.70)
life c-let. The duration of each larval stage and measurements
of body length were calculated in the manner previously discussed for
the other twro species and are included in Table 3.
"he adult stage was reached by the females
and by the males in an average of 175 brs.
in an average of 190 hrs
Roltnc.Th~e molting process of B. hanosnious was essentially
identical to that described For B. pa~ISsalus.
'Pn ndue:-in. The behavorial responses of 2. hameso3icus males
were observed to be almost identical to those exhibited cy males of
B. -a~ssl~us. Virgin females we~re approached directly even when obstacles
intervened and in several instances B. hemospicus males penetrated large
masses of P. redivivus specimens in locating and copulating with
conspecif'ic Females (Fig. 11a).
The process of copulation was intermediate between that described
for the twro other species. The spicules were thrust deeply into the
female, pernatracing~ the entire depth of the vagina (Fi,. 11b). Th~e
guberraculum was not inserted into the vagina and the spicules were forced
into the enfolding ring of the gubernaculum to the expanded upper
portion of the calomus (Fig. 1'b). During copulationi, m~uscular action
of the female genitals were observed similar to that described for
E. ecius.The spicules were pulled toward the single ovary, not
toward the post-uterine sack in which sperm were later observed. The
laminee of the spicules of this species are much more stoutly const_,cted
than those of B. lacinus, however, and did not bend. The ce halated
heads of the spicules and gubernaculum turned within the male's body
(Fi. 1c).Reproductive isolation of B. hremospicus was confirmed by
the rep;'odu~ctive isolation tests.
ceedine.~ The predaceous habits of B. ham~osicuis were similar to
those described for the other two species. Figure 12 shows an example
of cannibalism in an underfed, overpopulated colony, under which condi-
tion~s cannibalism was commrronly observed. The list of prey for the two
preceding species is identical with that of B. han~osnisus.
Competition studies resulted in ,he eventual annihilation of B.
h nnscicas populations under all mixed colony situations except in
colniztion with B. nassalus in which case the smaller species, B.
Mr ~s-lcus prevailed.
Fig. 11. Copulation of Eutle--ius iamo~spic-is.
a. The male penetrated this mass of prey
nie;.atodes In c dier to mate (arrow).
b. Spicule penetration, note position of
gu'oernaculum and capitulumn. c. Rotation
of spicules and gubernaculum due to female's
Fig. '2. Cannibalism by Eutlerius hamospicus.
Note material in lumen of the oesophagus of'
attacking specimens and dilated oesophageal
lumrren of the attacked male.
Stage E.S7.. ""iu .Tasls 3 am ei-:F
Oviposition in 1-cell 0 0 0
2-cell .5 :.0 1
4-cell '.5 3. .0
8-cell 3.0 6.0 5-5
16-cell 6.5 12.0 '3;
32-cell 7.5 15.0 i.
Morula 9.0 15.0 17.25
Blastula 13.0 21.0 15
Gastrula 15.0 26.0 ."
Tadpole ;6.0 30.0 ;".75
1st Larval 18.0 jo0 .C
Intrachorionic Molt 27. 0 wE.) 6.
2nd Larval 4-2.0 ?4.,0 6
Molt 3rd. Larval cs 00.0 109i.0 .25
9 67.0 115.0 9.
Molt 4th Larval 81.0 15. 35.~
$ 84.0 159.0 150.3
Molt Adult C 102.0 203.0 7.
Studies on reproductive h~abits of each of the -cl;ee species of
Bu:1'erius demonstrated that the males can locate iel;.ales or` theiCr on;;.
Three new species of the gen~us 31 .Oiy 99 r
found in Florida and the taxonom~ic diescriptionswere;~ ;-pirepar. The.
names Butlerius lacinus, B. cms-slus and B. hamOSrios er ss Ld
The embryonic and larval development of' each species was studied. Th
average time lapse per d~evelopm;ental ;tag f or each species is given in
Table Lc. Average Tim~e Lapse of th~e Developm~ental
Buteris sp. in rs from oviposition
S;Eeze~s fromn distan;ces of upj to 80 mm1 under in. vitr; ccr~ditions,
despite inti-ecmeiate masses of prey nemratodes and other indiv-iduals of
th-eir owJn species. N~one mated with individuals of' the other two species.
An indication of a~ccumulation of experience was exchibited in the cop-
ulatory responses of previously mated males.
The predaceous habits of the three species were similar. The
cuticle of large prey nematodes was drawn into the stoma of specimens
of Butlerius by suction. This suction resulted from -,he repeated, rapid
elongation of the stoma. The cuticle was then ruptured by the dorsal
tooth and the body contents ingested. Small prey nematodes were ingested
in1 too. 7 Dulring the latter type of feeding the dorsal tooth was writh-
draw~n into the wall of the stoma.
None of the three species of Bult'.e~ius described was as competitive
as iiYonenrchoides sp. or Tor-laimes suibtilis. Butlerius l~acinus was more
competitive than~r the o-ther two Bu~tlerius spp., whereas B. passalus w~as
the least comp~etitive.
1. Adam, W. 193C. Ein neuer freile~~cnce Namatone aus car E~c_:
Butlezi-a fiicu ,,s n. sec. Zool. ,LnTz. LeIOy C/1:139-142.
2. Cobb, N. A. 191'7. The Monor.ch;s. Soil Science. 3(5):--:2?.
. Chitwood, B. G., and M. B. ChitwooG 1938. Irureduction to r.,;ma-
tology. Baltimore 2;3 pp.
c;. Goodley, J. B. 1957. Labdora~tory nethod~s 'or worrke with plant and
soil nemratodes. Tc.Bl.Mns.ArLn.2
5. Goodey, T. 1929. On. some new~k an-d little-known~ free-livin~g n~em-a-
todes. J. Helminthol. 7:i7-62.
6. Goodey, T. 1951. Soil an;d freshwrater rnematodes. Xetheunr, Lon-d;-.
7. Goodey, T. ;963. Soil and freshwater r~nematodes. 2nu~ Ed. Edited by
J. B. Goodey. Methuen, London. 5 4 pP-
8. Lordello, L. G. D., and A. P. L. Zamith 1959. Obje;vac~es s'dore
0 g~nero "Butlerius" de n~emrat~deos de vida livre. R~ev. Biras.
9. Meyl, A. H. 1957. Two new 're~-;living, ;-.s~atodes, found i.- the
rain-water reserve of' Ques-eli 3s(V1. z (rmla
ceae) from Brasil. Pr-oc. Helminth~-~ol. Soc. Wa:sh. 24:j2-66.
10. Meyl, A. H. 1960. "lFrelebenden huematode, n" IL: Di iew
Mittieleuropa~s, edited by P. B-rokera;, ". Ehrmann, ana I;. Cla;-e-
1(5a):164 pjp. ;eipzia, Quelle &- ieyer.
11. Rahm, G. 1938. Freilebende und saprop~hytisch~te :l~;~oCatoCe dz~ r ste
Hainan. Anot. Zool. Jap. 17.046-067.
12. Russell, C. C. 1964. A rearing medlica for microphagous comatedes."~
The Florida Entomologist. L47(4):263.
13. Schuu:rmans-Stekhoven, J. H., and R. J. Hi. Teunissen 1938. Ne:;tod-s
liores terrestr-es. Exp~lor. Parc=. Nuat. A;Lbers (MIlssio -913-35),
Brussels 229 pp.
14C. Taylor, D. P. 19614. Butle~-- 1e re-~---:.:-. (Nea~toms, C13logistel~ eri
a new species of predaceous nematode from Illincia. Proc.
Helminthol. Soc. Wash. 31(2):129-132
15. Thorne, G., 1961. Principles of nematology. McGraw-Hil ,
New York. 553 PP.
C:;arles Clayton Russell was born October 9, 1937, at Key West,
Florida. He attended public schools in Sanford, Florida and was
graduated in 1955 fromn Seminole High School in Sanford, Florida. He
attended Pieomont College from 1955 until 1957 when he transferred to
the Un~riversity of Florida. He received the B~achelor of Science in
Agriculture degree in June, 1960 and the Master of Science in Ag-riculture
degree in Decemb~er, 1962.
He is a member of: Sigma Xi, Phi Sigma, Gamma Sigma Delta, Alpha
Zeta, Society of' Nematologists, Florida Nematology Forum, Newell
Entomological Society and Florida Entomological Society.
He is married to the former Carole Ann Bridges and they have a
son, John Charles, and a daughter, Cynthia Anin.
This dissertaion w~as prepared under the direction of the chairman
of the candidate's supervisor-y committee and has been approved by all
nembers of that committee. It was submit sted to the Dean of the College
of Agriculture and to the Graduate Council, and was approved as partial
fulfillment of the requirements for the degree of Doctor of Philosophy.
August 12, 1967
Lean, Zollege of Agriculture
Dean, Graduate School