Journal of Applied Animal Research

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Phylogenetic Taxonomy of Rumen Ciliate Protozoa

Based on Their Morphology and Distribution

Soichi Imai

To cite this article: Soichi Imai (1998) Phylogenetic Taxonomy of Rumen Ciliate Protozoa Based
on Their Morphology and Distribution, Journal of Applied Animal Research, 13:1-2, 17-36, DOI:
10.1080/09712119.1998.9706670

To link to this article: https://doi.org/10.1080/09712119.1998.9706670

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J. Appl. h i m . R ~ s 13
. (1998): 17-36

Phylogenetic Taxonomy of Rumen Ciliate Protozoa

Based on Their Morphology and Distribution
Soichi Imai'

Department of Veterinary Parasitoloky

Nippon Veterinary and Animal Science University
Musashino-shi, Tokyo 180,Japan
(Received October 9,1997; accepted December 23,1997)

Abstract
Imai, S. 1998. Phylogenetic taxonomy of rumen ciliate protozoa based
on their morphology and distribution. J . Appl. h i m . Res., 13: 17-36.
The ciliates belonging to the family Ophryoscdecidae in the suborder
Entodiniornorphina peculiarly inhabit the rurnen of various
m minants. They possess specialized and complicated bodies which
seem to have evolved with the evolutlon o f their hosts. Comparative
morpho1og.v o f these ciliates would be interesting as a model of
t.o-evolution between these parasites and their hosts. As a msult of
morphologic cnmparison of 14 genera o f this family, a temporary
phylogenetic tree containing a line connected continuously from
simpler species to more complicated ones, and a line independently
derived on the way from the genus Diplodinium can be created
However, there is no evidence that the morphologic relation of
ophryostnlecid genera exprcssrs the correct phylogenetic relation of this
family. Thus it is necessary to examine this problem by another
appmac.h. When the relation o f the distribution of respective genera of
Ophryoscolecidae, in which the geographic factor is excluded as f a r as
possible, to an advanced phylogenetic tree of host mammals is
examined, we can draw another temporary phyhgenetic tree. As
compared to this Fgure with the morphologic relation of ophryoscolecid
ciliates, the phylogenetic relation o f these ciliates seems to agree as a
I'el +81 422 31 4151, fax +81 422 31 4139; e-mail: LDP03073k3niftyserve.or.jp.
17
J Appl Anin] Res 0971-2119/98/$05 OOOGSP, India
18 Soichi Zrnai

rule. Accumulation of more detailed ahta from various hosts and their
careful comparison would be necessary in future. In such examinations
the species which are interesting phylogenetically may be discovered
from unemmined animals, especially wild ones.
Key words: Ciliate, Ophryoscolecidae, phylogeny, rumen protozoa,
ruminant, taxonomy.

Introduction
The rumen of ruminants, which are major large herbivorous mammals, has
superior morphology and function to degrade fiber of plants efficiently. In
general, herbivorous animals have to strive for promotion of fiber degradation
by retaining it in their intestine for long time, because fiber of plants, such as
cellulose, pectin and lignin are difficult to decompose. For this purpose, a part
of intestine of most herbivores expands and the plant materials are stored
there. Since all the herbivores have no cellulolytic activity of their own, they
must entrust fiber degradation to microorganisms inhabiting their digestive
tract. Some herbivores have expanded caecum and/or colon (hindgut
fermenter), and the others have large forestomach (foregut fermenter) such as
ruminants. The enlargement of foregut has various merits such as direct
utilization of microorganisms as nutrients, relatively easy control of microbial
ecosystem by saliva as buffering system and easy transmission of microbes to
neonates. Thus the ruminants and microorganisms inhabiting their m e n
have established a kind of symbiotic relationship during the long process of
evolution. Rumen microorganisms comprise viruses, bacteria, fungi and
protozoa (Ogimoto and Imai, i981). Of them, protozoa have larger bodies than
the others and characteristic shape which is easily observed by light
microscopy. facilitating their examinations which have performed up to this
time. Most of these protozoa are ciliates which are peculiar in the digestive
tract of mminants and can be classified into relatively small taxa. These
symbionts and their hosts might be interesting as a model of co-evolution.

Classi ficat ion o f r umen ciliates

Over 250 ciliate species have been described from various ruminants
(Williams and Coleman, 1991). They can be divided into two orders in the
class Trichostomatida, Vestibuliferida and Entodiniomorphida (Small and
Lynn, 1985), the species of which are described from the digestive tract of
various vertebrates, and almost all the members of Entodiniormophida
inhabit rumen or large intestine of large herbivorous mammals. The ciliate
density in respective habitats in generally very high, which ranges between
 Phylogeny of lumen ciliate protozoa 19

lo5-lo6/ ml of intestinal contents (Ogimoto and Imai, 1981). Members of the

order Entodiniomorphida consisting of 9 families of 3 suborders have more
complicated body structures than those in the order Vestibuliferida, epecially
ciliates belonging to the subfamily Entodiniomorphida have unique body
composition (Fig.1).
The entodiniomorphid ciliates detected from rumen consist of 3 families,
Orphpscolecidae in the suborder Entodiniomorphina, Blephamcorythidae in

l3.g. I . Ophryoscokx oaudatus. One of the most cnmplicated species in the

rumen ciliates, which is relatively cnmmon species in sheep. SEM
phntomicmgraph. Rnr = 50 pm.

the suborder Blepharocorythina and Buetschliidae in the suborder

Archistomatina. Of them, ophryoscolecid species are always the most
predominant in the rumen, which generally account for over 8@Yo of all the
rumen ciliates. The ciliates of Blephamcorythidae and Buetschliidae are in
low density in rumen but are predominantly detected from caecum and colon
of hindgut fermenters such as horse. Although the ciliates belonging to the
family Isotrichidae of the order Vestibuliferida are also found in rumen, both
the species diversity and density are low. Thus, ophryoscolecid ciliates would
be the best group to discuss on the co-evolution between rumen ciliates and
their hosts.
Generic and species classification of the family Ophryoscolecidae was
fundamentally performed by the monograph of Dogiel (1927).This monograph
is still useful for the identification of rumen ciliates, however, there are some
problems in the description of generic names, causing taxonomical confusion.
Dogiel (1927) divided this family into 6 genera and 6 subgenera. Slightly
20 Sotc*hi Imai

later, Kofoid and MacLennan (1930, 1932, 1933), who examined Lumen ciliate
compositions of Indian zebu, readjusted Dogiel's classification. In their
description, all the subgenera established by Dogiel were raised to genera and
14 genera were created based on the number and situation of contractile
vacuoles in addition to former characters for classification. Although most of
American and English researchers supported their classification, the
investigators in France opposed their stand, and Noirot-Timothee (1960)
supported establishment of subgenera by Dogiel. Although these two types of
classifications of Ophryoscolecidae are still used by some investigators,
revised classification advocated by Latteur (1966) seems to be more
reasonable. He adopted only the number and shape of skeletal plates as the
criteria for the Diplodinium group and contended to delete 3 genera,
Eodinium, Diploplastron and Eremoplastmn, from the 14 genera established
by Kofoid and MacLennan. However, Imai (1995) claimed to leave Eodinium,
which was put in the genus Diplodinium by Latteur, as a n independent
genus, because this genus possesses plural species, rod-like macronucleus
unlike that of Diplodinium and unique position of posterior contractile
vacuole. The subgenus Amphacanthus of the genus Entodinium described by
Dogiel (1926), which is peculiarly found from camels was cleared to be
homonym, so that it was redescribed as an independent genus
Campylodinium by Corliss (1978). However, from the findings that no
difference was cleared except the shape of processes of ectoplasm by Imai and
Guirung (1990), it was decided that this genus should be treated as a
synonym of the genus Entodinium. The taxonomy of the family
Ophryoscolecidae in generic level based on these findings is shown in Table 1.

Techniques for the identification o f rumen ciliates

For fixation and staining of rumen ciliates, the use of MFS solution is the
simplest method. MFS (methyl green-formalin-saline) solution consists of the
following components and looks dark green.
35 o/o formaldehyde solution (formalin) 100 ml
Distilled water 900 ml
Methylgreen 0.6 g
NaCl 8.0 g
Mix well, seal up tightly and store in a dark place. One or two liter of the
solution can be made beforehand and stocked. If the solution is exposed to
light, methylgreen dissolves into methylviolet which stains the ciliate bodies
 Table 1
The taxa of rumen ophryoscolecid protozoa and included genera
Phyl uni Ciliophora Doflein, 1901
Subphyluni Rhabdophora Small, 1976
Class Lilostoniatea Sniall & Lynn, 1981
Subclass Trichostoniatia Butschli, 1889
Order Entodiniomorphida Reichenow in Doflein & Reichenow, 1929
Suborder Entodiniomorphina Reichenow in Doflein & Reichenow, 1929
Family Ophryoscolecidae Stein, 1869
Subfamily Entodiniinae Lubinsky, 1957
Genus Entodinium Stein, 1859
Subfamily Diplodiniinae Lubinsky, 1957
Genus Diplrdiniurn Schuberg, 1888
Genus Eodiniurn Kofoid & Maclennan, 1932
Genus Eudiplodiniurn Dogiel, 1927
Genus Ostracodiniurn Dogiel, 1927
Genus Enoploplastmn Kofoid & MacLennan. 1932
Genus Metadiniurn Awerinzew & Mutafowa, 1914
Genus E1ytn)plastmn Kofoid & MacLennan, 1932
Genus Polyplastron Dogiel, 1927
Subfaniily Ophryoscolecinae Lubinsky, 1957
Genus Epidinium Crawley, 1923
Genus Epiphstmn Kofoid & MacLennan, 1933
Genus Opisthotrichurn Buisson, 1923
Genus Ophryoscokx Stein, 1859
Genus Caloscokx Dogiel, 1926
to violet, and later on ciliates are discolored. Samples should be added to 5-10
times the volumes of MFS solution, then only the nuclei of ciliates are
stained. Specimens fixed with MFS solution, preserved in glass tubes with
screw caps and stored in a dark place can be kept in good condition for 2-3
years, however, observation as early as possible is recommended. If these
samples are packed into small non-glass tubes capped tightly, they can be
sent by mail. Although MFS solution itself can also be kept for long time, it is
preferable to remake within one year interval.
General formalin solution (Ca 3% formaldehyde aqueous solution) is also
used as fixation agent. For the observation of large species containing many
food particles and fragments of plants in their endoplasm, the fixative
containing l/1 volume of glycerol is often used to examine skeletal plates.
General methods for histopathological examinations such as
haematoxylin-eosin staining and silver-impregnation methods are also
22 Soichi Zmai

available for rumen ciliates (Wolska, 1966; Ogimoto and Imai, 1981;
Fernandez-Galiano et al., 1985). For the scanning and transmission electron
microscopy, other articles eg. Ogimoto and Imai (1981) and Williams and
Coleman (1991)are recommended.

Morphology and criteria of ophryoscolecid ciliates

The cells of the family Ophryoscolecidae have generally flattened ovoidal body
(Fig. 2). Ciliary apparatuses are restricted on the peristome and its

Fig. 2-5 :SEM photomicmgmphs of ophryo.scolecld ciliates (Entodinium spp.).

2. Whole hody of Entodinium sp., Cilia are restricted amu.nd the
pcristomal zone, which is diuicied into two groups, inner (ZAZ) and
o u k r (OAZ) adorn1 zones. The surface without cilia is striated.
Rar=20 p m 3. Cilia of outer adoral zone forming closely packed
hundles ,(arrow). Rar=5 p m . 4. Retmcted ciliary zone fractured
lcngthiuise. Adoral cilia (AC) are put in a triangular chamber, and
adoral lips (AT,) close the exit of the cilia. Rar=20 pm $5. Surfam
fractured C I C ~ S Sthe right-kft sides of a ciliate with extending adoral
cilia. Cilia arc extending along the wall of deep uestibulum (armiu).
Rar=lOum.
 Phylogeny of rumen ciliatepmtozoa 23

neighboring area at the anterior part of body and the other surface is covered
with relatively thick pellicle without cilia. Pellicle is often highly developed
and forms caudal processes at the posterior part of the body. The surface of
pellicle has species specific striations (Imai and Tsunoda, 1972; Imai et al.,
1977). Cilia form closely packed groups named syncilia (Fig. 3). Ciliates
locomote using these cilia while they revolve. Ciliary zones can be retracted
when the circumstance becomes adverse (Fig. 4). Adoral ciliary zone can be
divided into two, outer and inner, zones (Imai et al., 1983) and extend to the
cytostome along the wall of relatively deep vestibulum (Fig. 5). Cell is divided
into endoplasm and ectoplasm. Endoplasm is relatively narrow and consists of
cytosol and many food vacuoles including starch grains, fragments of plants
and bacteria. Accordingly it appears to be just like a stomach in polycellular
animals. Ectoplasm is thick and contains one or more contractile vacuoles and
many amylopectin granules like rice grains (Nakai and Imai, 1989) as reserve
food materials. A cytopyge is present at posterior end of the body and short
rectum extends towards endoplasm. A macronucleus is rod-like in general,
but sometimes shows more complicated aspects in compliance with species.
Micronucleus is one in number except during binary fission and situated
besides the macronucleus (Fig. 6). Most of the large species in this family
have skeletal plates consisting of assembly of polysaccharide pieces in the
ectoplasm. Skeletal plate is assumed to be supportive structure rather than
storage one.
Of these structures, the number and situation of ciliary zones, shape of
macronucleus, the numbers and situation of contractile vacuoles and the
number and shape of skeletal plates have been recognized to be stable
characters, so that these characters are used as criteria for the identification
of genera and species of the family Orphryoscolecidae. As the classification of
genera, num3er and situation of ciliary zones, number of contractile vacuoles,
number and shape of skeletal plates and shape of macronucleus can be used
as criteria for identification (Figs. 7, 8). For species identification, shape of
total body, especially anterior end, shape and length of macronucleus and
number and situation of contractile vacuoles are the criteria. Body size is also
often in use. Caudal spines at the posterior end of body had also been used as
a criterion (Kofoid and MacLennan, 1930, 1932, 1933) for species
identification. However, now this character is not used at least as a species
criterion, because it became clear that caudal spines easily change by the
condition of circumstances from the results of in uitro culture (Coleman, 1979)
and transfaunation between different hosts (Imai et al.. 1994).
24 Solchi I m a i

Fig. 6 : Imgitu.dina1 sectiori of Pidyphstmn multiucsic.ulatum frcyuoitly

/ ? w i d from sheep and cattle. Thick cvtopiasm (Ec) arid cndoplnsm
(En) with man,y food uacuoles cnntaining starch grains (SG) and
bacteria ( R ) art’ distinct. Adoral ciliary zone (ACZ), mncronuclcus
(Ma), micmnucleus (Mi) and cytopyge (CP) art’ also shoiun. Rnr=.5O
Clm.
Species identification of large ophryoscolecids such as the genera,
Diplodinium, Eudiplodin.ium and Ostmmdiniurn is relatively easy, but that
for small and no caudal spinated entodinia is fairly difficult.

Rumen as a habitat of ciliates

It is easy to imagine the reason why rumen ciliates and the other
microorganisms adopted the gastrointestinal tracts of the hosts as their
habitat, because this place guarantees very stable environment made by the
homeostasis of the host and supply of food taken by the host periodically.
Since most ciliates can obtain energy necessary for their life without aerobic
 Phy1ogen.y of lumen ciliateprotozoa 25

Fig 7 : Schematic drawing of Polyphstron multioesicuhtum. Various

structures in the h r e are used for the identification of genera and
species. ACZ. adoral ciliary zone; CP. cytopyge; CV, contractile
uacuole; LCZ. lefi ciliary zone; Ma. macronuclew; Mi. micronucleus;
OP opemulu m; R. rectum; SP. skeletal phte.

respiration, it is considered that the gastrointestinal tract of mammals was

suitable habitat for these facultative or strict anaerobic protozoa.
The ciliate species belonging to the suborder Entodiniomorphina
involving Ophryoscolecidae are found in the intestine of all the large
herbivorous animals such as Artiodactyla, Perissodactyla and Proboscida
(Ogimoto and Imai, 1981), and have no habitat other than gastro-intestinal
tract. From the fact, it can be assumed that the ancestors of these ciliates had
already inhabited the intestine of herbivorous animals in fairly early period
after the establishment of their hosts on the earth.
Of these 7 families consisting of the suborder Entodiniomorphina, the
family Cycloposthiidae predominantly found from the caecum and colon of
26 Soichi Zmai

Fig. 8 : Members of the family Ophryoscolecidae. a d Entodinium spp. One

ciliary zone, one contractile uacuok and rod-like macmnucleus. a. E.
simplex; b. E. himastus; c. E. tricostatum; d E. caudatum. e.
Diplodinium anisacanthum. Two ciliary zones, two wntractile
uacu.oles, an operculum and bending macronucleus are present. f.
E0diniu.m postemvesicu,latu.m. Two ciliary zones, two mntractile
vacuoles, posterior one of which is located at just posterior to
macmnu.clcus, an oprou1u.m and md-like macmnucleus. g
Ostracodinium ymcile. This genus has two ciliary zones, two or more
contmctile uacu.oles, an operculum, relatively complicated
macmnucleus and one broad skeletal plate. h Enoploplastmn
trilorimtum. This genus resembles Ostmcodinium, but it has one
hmad and two slender skeletal plates on both sides of the bmad one.

horses has wide distribution range; it is also detected from hippopotamuses

(Artiodactyla), tapirs, rhinoceroses (Perissodactyla) and elephants
(Proboscida). In contrast, most of the other families have high host specificity,
viz. the family Ophryoscolecidae is found only from Ruminantia and
Tylopoda, the family Ditoxidae from Hippomorpha, and Polydiniellidae from
 Phylogeny of lumen ciliate protozoa 27

Fig. 8 contd.
i. Kudiplodinium mcy;ii. Two ciliary zones, two contratile vacuoles, an
opercu lu m, rolatively cnmplicated macronucleus and one slender skeletal plate.
j . Metadinium tauricum. This genus resem bles Eudiplodiniurn, but two
relatively large skeletal plates are present. k. Polyplastmn rnultivesciulatum.
Two ciliary zones, contractile vacuoles, a distinct operculurn, md-like
macrnndeus and two l a z e and three small skeletal plates situated on right
and left sid.w respectively. 1 Elytmplastron bubali. This genus is similar to
Pol.yplastmn but it has fou.r contractile vacuoles situated in tandem and three
large skeletal plates one of which is located the opposite sick to the other two.
m. Epidinium ecaudatu.m. Two ciliary zones, dorsal one of which is located
slightly posterior to adnral zone, two Lvntractile vacuoles, no operculum,
&-like macmnu~:kw.sand thnv skeletul plates at vcntml side. n. Qwisthotrichum
janus. This genus resem bles Epidiniu.m, but the dorsal ciliary zone is situated
fairly posterior to the adoral zone. 0. Ophryoscoles cau.dutu,s. Two ciliary zones,
dorsal one of which forms a girdle amund the middle of the body, nine
contractile vacuoles arranged in two transverse rows, no opercu.lum, md-like
macmnucleus, three skeletal plates at ventral side and many furcated spines at
posterior part of the body. p . Caloscolex came1inu.s. Two ciliary zones,
relatively large hdnral zone and dorsal zone forming a girdle around the
middk of the body, seven cnntractik vacuoles arranged in transverse mw, no
operculum, rod-like macmnu.cltw,s and three large skeletal plates forming a
fan-like plate. Each har=30 pm.
28 Soichi Imai

Table 2
Distribution of rumen ophryoscolecid genera in various hosts*

Camel (Camelus) + + + + +
Lama (Lama) + + +
Mouse deer (Tragulus) t + + +
Muntjac (Muntiacus) t t
Fallow deer (Duma) t t + +
Red deer (Ceruus) + t t + t t
Japnese deer (Ceruus) t + t +
Water deer (Rusa) t t
White-tailed deer t
(Odocoikus)
Moose (Alces) + + + +
Reindeer (Rangifer) + + + + + + t + +
Roe deer (Capredus) +
Giraffe (GimHa) + t t + t +
Zebra duiker (Cephalophus) + t +
Water buck (Kobus) + + +
Reed buck (Rcdunca) + + +
Sable antelope (HippotmfCus) + + +
Sassaby (Damalisc-us) + + + + + +
Royal antelope (Ncotrugus) + + +
Lechewe (1n.c.h) t + + + +
Impara (Aepycems) + + + + t
Gazelle ( C k ~ e l l a ) +
Springbok (Antidomas) +
Dikdik (Madoqua) + + + + +
Steinbok (Raphicerus) + + t + +
Japanese serow (Capriconis) + + +
Chamois (Ru,picapra) + + t t t
Musk-ox (Ouibos) t + + + +
Kudu (Tmge1aphu.s) + + + + +
Waterbuffalo (Rubalus) + + + + + + t + + + +
Cattle, Zebu, Yak (Ros) + + + + + + + + + + + +
Af'rican buffalo (Syncrrus) + + t + t +
Bison (Bison) + + + t +
Goat, Ibex (Capra) + + + + + + + + + +
Sheep (Ouis) + + t t t t + + + + +
*&vised 0Kimot.u and lmai (1981).
 Phylogeny of lumen ciliate protozoa 29

Elephantoidea (Ogimoto and Imai, 1981). These findings suggest that the
ancestor of cycloposthiid ciliates already established in the intestine of
common ancestor of large herbivorous mammals such as Artiodactyla and
Perissodactyla. Consequently it is difficult to consider that ophryoscolecid
ciliates evolved from free-living ciliates after the period when ruminants
established. Ruminants have developed as the enlargement of grassland
consisted of monocotyledon from the Miocene epoch, Cenozoic era, and a t
present they have become firm as the major large herbivorous mammals
around the world. Thus we can imagine that ophryoscolecid ciliates have
evolved with the evolution of their hosts in the isolated environment, if
assuming that the ciliates belonging to the suborder Entodiniomorphina
already established in the rumen when ruminants appeared on the earth.

Distribution of ophryoscolecid ciliates

When the distribution of respective ophryoscolecid species was examined on
the various ruminants inhabiting the present world, these are divided into
two types, the species showing wide host range and those found only in
limited hosts. Ciliate composition rate of respective types in the rumen is
much higher in the species having wide distribution than those with limited
host range.
Most of the protozoan parasites inhabiting the intestine of the hosts
generally form resistant cysts in the posterior part of intestine and those cysts
excreted in the feces become new source of infection. Rumen ciliates inhabit
anterior part of intestine of hosts, so that both trophozoites and cysts are not
detected in the feces, because these are decomposed by digestive enzymes
secreted from the small intestine of the hosts (Imai et al., 1981; Kurihara and
Katsuno, 1964). Accordingly infection of the ciliates to the other hosts,
especially to newly born neonates has been considered to occur
mouth-to-mouth oral transmission via saliva and ration which include the
ciliates sent from rumen to mouth by rumination behavior of hosts (Mangold
and Radeff, 1930). The ciliate species inhabiting high composition rate in the
rumen have more chances to be sent to mouth with saliva, therefore these
ciliates have more chances to infect other hosts and show wide distribution
range. However, infection occurs only a t close range as between mother and
child, and difficult to occur a t f a r distance, since the ciliates cannot live for a
long time outside the rumen. For this reason, it is expected that the ciliate
species with low compositon rate infect a t most among the host individuals in
a small group and raise their host specificity. Most of the wild ruminants form
homogeneous groups for their life and exclude the invasion of the other
30 Soichi Imai

Fig. 9 : Tentative relationship among respective genera in t h e frmily

Ophqoscolecidue hascd on their morphologic chamcters. a-e.
Entodinium, f. Diplodincum, g. Eodiniurn, h Ostracodiniurn, i.
Enoplopla.str~iri,j . Eudiplodinium, k Metadiniurn, 1. Elytroplastron,
m, Polplastn~ri,n. Epidiriium, 0. Opisthotrichu rn, p . Epiplastmn, q.
Ophryoscolcs, r. Caloscokx.
species of ruminants. In contrast, ciliate infection in domestic ruminants
seems to occur easier than in wild animals, because they are often fed
artificially in large number in a narrow area.
Although the host specificity of rumen ciliates is very high between
ruminants and the other herbivorous animals, infection of most rumen ciliate
species occurs among ruminants (Imai et al., 1994).Therefore, the infection of
most ciliate species seems to occur easily between the different species of the
hosts when they are kept close to each other. In contrast, each ciliate
composition is affected independently from the ciliate fauna of different
species of hosts inhabiting neighbouring area, so that different ciliate
compositions will be established even the hosts are of the same species, if they
are geographically separated.
When the ciliate composition of wild and domestic ruminants reported
so f a r are compared, the composition of wild animals is generally simple in
contrast to that of domestic animals which is plentiful (Table 2), suggesting
that the number of chances of infection strongly affected the formation of
ciliate compositon.
 Phylogeny of rumen ciliate protozoa 31

Phylogenetic aspects based on the morphology and

distribution of rumen ciliates
Based on the distribution of rumen ciliates in various hosts shown in Table 2,
the genera Entodinium, Diplodinium, Ostracodinium and Epidinium have
wide distribution range. As mentioned above, rumen ciliate composition in the
host seems to be determined by the phylogenetic factor in which the ciliates
are succeeded mainly from mother to her children and the geographic factor
which is decided by the number of chances for infection from various species
of hosts. Since above four genera are also found from various wild ruminants
which are generally isolated from groups of the other host species, we can
suppose that these four genera are phylogenetically ancient group. If so, it is
suggested that these genera have simple and primitive body structures.
Actually the genus Entodiniu r n has the simplest morphologic characters in
the ophryoscolecid group (Fig. 8), that is, the species in this genus have
generally small body, one ciliary band, one contractile vacuole, and no skeletal
plate. Although the genus Diplodinium has somewhat more complicated body,
larger body size, two ciliary bands, opperculum between them and two
contractile vacuoles, than those in Entodinium, the species of this genus have
no skeletal plate, showing that this genus is the second simplest group
following the genus Entodinium. In additon to these morphological
characters, these two genera have exceptionally many species respectively,
compared to the other genera. These facts suggest that these two genera are
prototype of Ophryoscolecidae. The genera Ostmcodiniurn and Epidinium
have more complicated bodies than those of above two genera. Ostracodinid
species have one wide skeletal plate a t the right side of the body in addition to
the structures found in the diplodinid species. On the other hand, the
morphologic characters of the genus Epidiniurn are fairly different from these
three genera, that is, epidinid species have slender body, no opperculum,
dorsal ciliary band situated a t posterior part to adoral ciliary band and
complicated skeletal plates.
From these findings, temporary phylogenetic lines can be created
containing a line connected continuously from simpler species to more
complicated ones and a line independently derived from the genus
Diplodiniu n. Both Ostmcodiniurn and Epidinium have several related
genera showing more complicated body, respectively. The genera Metadinium,
Elytroplastmn, Enoploplustron and Polyplastron are related to Ostmmdinium
and the genera Opisthotrichum, Ophryoscolex and Calnscolex have near
relation to Epidiniurn. In these genera, there is a relation between the
morphology and distribution, in which more complicated species have more
32 Soichi Imai

Fig. I0 : A tcntatioc m&l 01 thc phylogenetk relation between the lumen

ophryoscolecid genera and their hosts. In this figure, the periods when
thp rvspctioe genera had deriocd from the ancestors of the present
hosts arc surmised (modified f m m D o g d , 1.947; Ogimoto and Imai,
1981).
 Phyhgeny of rumen ciliate protozoa 33

limited distribution range. A phylogenetic relation of these genera based on

their morphologic complicacy from simpler genus to more complicated one is
shown in Fig. 9.

Phylogenetic correlation betwen rumen ciliate

composition and host ruminants
There is naturally no evidence that the morphologic relation of ophryoscolecid
genera shown in Fig. 9 expresses the correct phylogenetic tree of the family.
Thus it is necessary to examine this problem by another approach. Since the
distribution range of ophryoscolecid species is almost limited in the rumen of
ruminants with only exception of the caecum of capybara (Dehority, 1987;
Imai et al., 1997), it can be assumed that these ciliates have co-evolved with
the evolution of ruminants. The evolution of ruminants has been well
examined based on the comparative morphology and fossils. Accordingly,
when the relation of the distribution of respective genera of Ophryoscolecidae,
in which the geographic factor is excluded as far as possible, to the
phylogenetic tree of hosts is examined, we can draw a figure as shown in Fig.
10. A s seen from the comparison of this figure with the morphologic relation
of ophryoscolecid ciliates shown in Fig. 9, the phylogenetic relation of these
ciliates seems to a k n e as a rule. However, the positions of several genera
such as Polyplastmn and Eudiplodinium which have wide distribution range
are still insufficient, because these genera have high possibility to infect
secondarily from the other host species. Accumulation of more detailed data
and careful comparison of these would be necessary in the future.
As mentioned above various groups of the ciliates belonging to the
suborder Entodiniomorphina are peculiarly found from the gastrointestinal
tract of many large herbivorous mammals. Phylogenetic approach of rumen
ciliate protozoa as performed by the author and his colleagues would be able
to extend to the other ciliate groups of this suborder. For example, the ciliate
species which is considered to be intermediate type between Ophryoscolecidae
and Cycloposthiidae has been described from the stomach of hippopotamus
(Thurston and Noirot-Timothee, 1973). In future, the species which is
interesting phylogenetically may be discovered from unexamined animals
especially wild ones.
Entodiniomorphid ciliates are the group which would not have appeared
if large herbivorous mammals did not evolve. When herbivores ingested a
large amount of plants which are difficult to digest, those foods might be
accumulated for relatively long time in their gastrointestinal tract.
Establishment of cellulolytic microbes which invaded with plants and water
34 Soichi Imai

would easily occur a t that time. Free living ciliates inhabiting the
environment with low oxygen and rich organic substances such as a ditch
would also be possible to establish in the digestive tract. If so, large
herbivorous dinosaurs such as Triceratops which had body similar to cattle
and had lived for long time by f a r than the present herbivores might have
such ciliates in their intestine.
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