Parasitol Res (2007) 100:855–860
DOI 10.1007/s00436-006-0346-1
ORIGINAL PAPER
Thelazia rhodesi (Spirurida, Thelaziidae), bovine eyeworm:
morphological study by scanning electron microscopy
Soraya Naem
Received: 3 September 2006 / Accepted: 13 September 2006 / Published online: 10 November 2006
# Springer-Verlag 2006
Abstract Thelazia rhodesi (Spirurida, Thelaziidae) is a
parasite of ruminants, which causes ocular infections.
Scanning electron microscopy was used to study the surface
ultrastructure of adult worms of this nematode. At the
anterior end of both sexes, the buccal opening was
orbicular. Around the mouth, four pairs of cephalic papillae
and two lateral amphids were seen. A pair of lateral cervical
papillae was present. In the female, the vulva was located at
the anterior part of the body, and the tail end was stumpy
with two phasmids near its extremity. In the male, the tail
was blunt, without caudal alae, and curved ventrally. There
were 14 paired preanal papillae, one single papilla directly
anterior to cloaca, one paired postanal papilla, and two
phasmids at the posterior end. The spicules were unequal
and dissimilar. The cuticle was transversally striated and
appeared serrated.
Introduction
Thelazia rhodesi Desmarest, 1822 is a parasite of domestic
and wild ruminants and less commonly of horses (Soulsby
1986; Anderson 2000). This nematode occurs on the
surface of the cornea, under the nictitating membrane, in
the conjunctival sac and lachrymal duct. The worms are
viviparous, and the first-stage larvae are passed by females
into the lachrymal secretions where they are ingested by
nonbiting Diptera. It has been confirmed that Musca
autumnalis and Musca larvipara are suitable intermediate
hosts (Anderson 2000; Giangaspero et al. 2004). Larval
S. Naem (*)
Department of Pathobiology, College of Veterinary Medicine,
Urmia University,
P.O. Box1177, Urmia, Iran
e-mail: sorayanaem@yahoo.com
development takes place in the thorax and abdomen of the
insect, and infective stages are present in 18–25 days.
Development to adult takes place without migration, and
the prepatent period is between 3 and 6 weeks. The first
stage of Thelazia is very short-lived in the lachrymal
secretion, only surviving a few hours, and transmission
depends upon the continuous presence of the vectors. For
this reason, thelaziasis has a seasonal occurrence according
to the seasonality of the intermediate hosts (Dunn 1978).
The adult and larval stages live in eyes causing conjunc-
tivitis, keratitis, lacrimation, ocular discharge, and ulcers
(Soulsby 1986; Otranto and Traversa 2005). Bovine
thelaziasis was first reported in Iran by Ebadi in 1951.
Other investigations indicate the presence of bovine
thelaziasis in Japan, Ghana, Afghanistan, USA, Canada,
and Italy (Okoshi and Kitano 1966; Vohradsky 1970; Barus
et al. 1976; Genden and Stoffolano 1980; O’Hara and
Kennedy 1991; Giangaspero et al. 2000).
Apart from the morphological descriptions (Kikuchi
1976; Guttekova 1987; Choi et al. 1989; Beelitz et al.
1997; Otranto et al. 2003a; Naem 2005) and molecular
findings (Otranto et al. 2001; Tarsitano et al. 2002; Otranto
et al. 2003b) that have already demonstrated the differences
among Thelazia species, no detailed morphological de-
scription of T. rhodesi has been presented. This article
reports most surface features of adult T. rhodesi by
scanning electron microscopy (SEM).
Materials and methods
From April to December 2002, 400 cattle were examined at
an abattoir located in Tehran City, Iran. The conjunctival
sac and lachrymal duct of both eyes were flushed with
sterile saline solution, and 20 adult worms were removed
856 Parasitol Res (2007) 100:855–860

from the eyes of eight naturally infected cattle. Seventeen Results

adult T. rhodesi (13 females and 4 males) were selected and
washed in 2% sodium cacodylate buffer (pH 7.2). The
nematodes were cleared with lactophenol solution (glycer-
in, lactic acid, phenol, and distilled water), and identifica-
tion of the species was confirmed on the basis of light
microscopic examination with reference to keys (Ershov
1956; Yamaguti 1961; Skrjabin et al. 1967; Skrjabin 1969;
Chabaud 1975; Lichtenfels 1975; Soulsby 1986). The
worms were next placed in 4% glutaraldehyde. The speci-
mens were then transferred from Iran to the EM Facility at
the Department of Pathology and Molecular Medicine,
McMaster University, Canada, for SEM study. The nema-
todes were put into fresh 4% glutaraldehyde for 2 h,
postfixed in 1% osmium tetraoxide in cacodylate buffer
(pH 7.2) at 4°C for 1 h, and dehydrated through a series of
increasing ethanol concentrations (50, 75, 96, and 100%).
They were dried with liquid CO2 in a critical point drier
apparatus (Critical Point Drier, Model # 2800, Ladd
Research Industries, Burlington, VT, USA). Each worm
was then placed on a numbered aluminum stab, which was
coated with gold in an ion-coating sputter (Sputter Coater,
Model # E5100, Polaron Instruments, USA) and then
viewed with a JEOL JSM-840 Scanning Electron Micro-
scope operated at 15 kV. Photographs were taken with a
digital acquisition system for analog SEMs (GW Electron-
ics, Norcross, GA, USA).
The cephalic region was similar in both sexes. The mouth
was orbicular and had no lips, with its anterior edge turned
back and cut into two festoons, starting with a small
capsule 10–14.5 μm wide at the buccal opening. Around
the mouth, four pairs of submedian small, conoidal cephalic
papillae and two lateral amphidal apertures were seen
(Fig. 1). Also, two lateral cervical papillae, one on each
side, 350–384 μm from the anterior end, were noticed
(Figs. 2, 3, and 5). At the anterior end of both sexes, an
excretory pore was seen (Fig. 4).
The females were 12.5–20.5 mm long and 300–500 μm
wide. At the anterior end of the body, the vulva was located
in the esophageal region, 505.2–536.3 μm from the
cephalic region (Figs. 5 and 6). The pattern of the cuticle
around the vulva was different (Fig. 6). Also, there was a
small papilla close to the vulva on the lateral side of the
female’s body (Figs. 7 and 8). At the posterior end, the anal
pore was seen (Fig. 9), and the tail end was stumpy with
two phasmids near its extremity (Figs. 9 and 10).
The cuticular surface showed coarse transverse stria-
tions in both sexes, giving them a serrated appearance
(Figs. 11 and 12). The bands of striation at the anterior,
middle, and posterior portions were 15, 20, and 30 μm
apart, respectively. The cuticle showed some tumor-like
masses at the posterior end of one female worm (Figs. 13
and 14).
The males were 7.5–14.5 mm long and 420–475 μm
wide. The tail was blunt, curved ventrally, and without

Fig. 1 Scanning electron mi-

crograph of the anterior end of
the male Thelazia rhodesi,
showing mouth, with its anterior
edge turned back and cut into
two festoons (arrows), cephalic
papillae (stars), and amphidal
apertures (A). Bar 10 μm
Fig. 2 Scanning electron
micrograph of the anterior end
of the male Thelazia rhodesi,
showing cervical papilla (CP).
Bar 100 μm
Fig. 3 Scanning electron mi-
crograph of anterior end of the
male Thelazia rhodesi, showing
a higher magnification of cervi-
cal papilla (CP) and cuticular
transverse striations (TS).
Bar 10 μm
Fig. 4 Scanning electron
micrograph of the anterior end
of the male Thelazia rhoesi,
showing excretory pore (EP)
and cuticular transverse
striations (TS). Bar 10 μm
Parasitol Res (2007) 100:855–860 857

Fig. 5 Scanning electron mi-

crograph of the anterior end of
the female Thelazia rhodesi,
showing cervical papillae (CP)
and vulva (V). Bar 100 μm
Fig. 6 Scanning electron micro-
graph of the female Thelazia
rhodesi, showing a higher mag-
nification of the vulva (V), cuticle
around the vulva (star), and
cuticular transverse striations
(TS). Bar 10 μm
Fig. 7 Scanning electron micro-
graph of the female Thelazia
rhodesi, showing cervical papilla
(arrowhead), vulva (V), and a
small papilla close to vulva (ar-
row). Bar 100 μm
Fig. 8 Scanning electron micro-
graph of the female Thelazia
rhodesi, showing a higher mag-
nification of a small papilla close
to the vulva (arrowhead) and
cuticular transverse striations
(TS). Bar 10 μm
Fig. 9 Scanning electron micro-
graph of the posterior end of the
female Thelazia rhodesi, show-
ing anal pore (AP) and phasmid
(Ph). Bar 10 μm
Fig. 10 Scanning electron mi-
crograph of the posterior end of
the female Thelazia rhodesi,
showing a higher magnification
of phasmids (Ph). Bar 10 μm

caudal alae, and the spicules were unequal and dissimilar
(Figs. 15 and 16). There were 14 paired preanal papillae,
one single papilla directly anterior to the cloaca, one paired
postanal papilla (Figs. 16, 17, and 18), and two nipple-
shaped phasmids at the posterior end (Figs. 17 and 18).
Discussion
This paper reports most surface features of adult T. rhodesi
(Spirurida, Thelaziidae) using SEM.
The cephalic characteristics of spiruroids present a
varied and complex picture. Different species belonging to
this genus are characterized according to the morphological
description of Yamaguti (1961). Among the Thelazia
species, Thelazia callipaeda and Thelazia californiensis
were known to cause human thelaziasis (Miyazaki 1991).
The arrangement and the number of the cephalic papillae in
T. californiensis appear to be the most generalized of any
known spiruroid of nematode (Chitwood and Wehr 1934).
In this study, T. rhodesi shows an internal circle of two
labial festoons, an external circle of four pairs of cephalic
papillae, and two lateral amphids. Gibbons (1986) also
showed the internal circle of labial papillae in Thelazia
gulosa. In Thelazia skrjabini, the head has two circles of
papillae: the inner circle with six papillae and the outer
circle with four papillae. Amphids are at both lateral sides
(Kikuchi 1976). T. californiensis shows an internal circle of
six very slightly reduced papillae, and an external circle of
eight well developed, symmetrically spaced papillae, each
papilla of the external circle being situated at the same level
on the head (Chitwood and Wehr 1934). Choi et al. (1989)
observed two large head papillae on the mouth opening of
the male T. callipaeda, which were absent in the females
and were distinct from the cephalic papillae in their
morphology and orientation. In another investigation on
T. callipaeda, several pairs of the nipple-shaped cephalic
papillae were reported (Arizono et al. 1976). SEM
observations of the female T. callipaeda showed only one
pocket-shaped amphid at the anterior end (Choi et al.
858 Parasitol Res (2007) 100:855–860

Fig. 11 Scanning electron

micrograph of the female
Thelazia rhodesi, showing
cuticular transverse striations
(TS). Bar 10 μm
Fig. 12 Scanning electron mi-
crograph of the posterior end of
the female Thelazia rhodesi,
showing cuticular transverse
striations (TS). Bar 10 μm
Fig. 13 Scanning electron
micrograph of the female
Thelazia rhodesi, showing
cuticular tumor-like masses
(arrows). Bar 100 μm
Fig. 14 Scanning electron mi-
crograph of the female Thelazia
rhodesi, showing a higher mag-
nification of cuticular tumor-like
masses (stars) and cuticular
transverse striations (TS). Bar
10 μm

1989), while Otranto et al. (2003a) observed two amphids
in lateral position and four submedian papillae on both
sexes of T. callipaeda. Nematode amphids exist in a variety
of forms and sizes, some probably being purely chemo-
sensory while some being photoreceptive, with an associ-
ated gland (McLaren 1974). The functional type of the
amphids of T. rhodesi is yet unknown. In this study, two
cervical papillae at the anterior end of the worm were seen.
These papillae were paired, simple, and similar in shape.
The cervical papillae are often very small and inconspicu-
ous, which has led to the erroneous conclusion that they are
absent in some species. The larger, conspicuous cervical
papillae vary in form from thin, needle-like appendages to
complex structures with denticulate posterior borders. The
position, size, and shape of the cervical papillae are used as
taxonomic characters. It is likely, in view of their structure

Fig. 15 Scanning electron micro-

graph of the posterior end of the
male Thelazia rhodesi, showing
spicules (S), precloacal papilla
(PrCP), postcloacal papilla (PoCP),
and phasmid (Ph). Bar 10 μm
Fig. 16 Scanning electron micro-
graph of the posterior end of the
male Thelazia rhodesi, showing
spicule (S), one single papilla di-
rectly anterior to cloaca (black
arrow), precloacal papilla (PrCP),
and postcloacal papilla (PoCP).
Bar 10 μm
Fig. 17 Scanning electron
micrograph of the posterior end
of the male Thelazia rhodesi,
showing precloacal papilla
(PrCP), cloaca (C), postcloacal
papilla (PoCP), and phasmid (Ph).
Bar 10 μm
Fig. 18 Scanning electron micro-
graph of the posterior end of the
male Thelazia rhodesi, showing
postcloacal papillae (PoCP) and
phasmids (Ph). Bar 10 μm
Parasitol Res (2007) 100:855–860
and position, that they function as mechanoreceptors
(Gibbons 1986). These organs determine whether the
nematode can pass through a small space (McLaren
1976).
Thelazia species have morphological differences such
as the location of the vaginal opening, the number of
cuticular transverse striations, and the number of caudal
papillae. The results of the present study indicate that the
vulva was located 505.2–536.3 μm from the anterior end
of T. rhodesi. The vulva of T. skrjabini was located 0.4–
0.7 mm from the anterior end and protruded (Kikuchi
1976). Another investigation showed that the vulva of
Thelazia lacrymalis was situated 593 μm from the anterior
end (Beelitz et al. 1997). In a study which was carried out
by Otranto et al. (2003a) on T. callipaeda, the vaginal
opening was located at 62.0–162.2 μm anterior to the
esophago–intestinal junction.
In this study, the cuticle had prominent transverse
striations in both sexes. An electron microscope study of
the cuticle of T. gulosa and T. rhodesi revealed very
interesting architectonics of the sculpture of the body surface
of these nematodes (Guttekova 1987). The surface layer of
the cuticle was ridged crosswise and lengthwise. While
certain uniformity of the shape and distribution of the surface
structures could be seen in T. gulosa, the case was contrary
in T. rhodesi (Guttekova 1987). The ridges and projections
of the cuticle had different structure and incidence in
particular thirds of the nematode body. The unusually
formed body surface structure found in Thelazia complied
with the fixation and locomotion of these nematodes at the
exacting place of their host’s eye (Guttekova 1987). In T.
skrjabini, transverse striae were not salient, and short
longitudinal lines were found on the cuticle. The length of
striae and the longitudinal lines were short at the cervical
part and longer at the midbody (Kikuchi 1976). The results
of the present study indicated bands of striation at the
anterior, middle, and posterior portions that were 15, 20, and
30 μm apart, respectively. The numbers of cuticular
transverse striations of T. callipaeda at the head, middle,
and tail portions were 400∼650, 250, and 300∼350 mm−1,
respectively (Choi et al. 1989). Another study, which was
carried out on T. lacrymalis by Beelitz et al. (1997), showed
that the numbers of cuticular transverse striations at the three
above-mentioned portions were 219, 287, and 267 mm−1,
respectively, in the male, and 215, 293, and 276 mm−1,
respectively, in the female worms.
Results of the present study indicate that the numbers of
caudal papillae of the male worm are different from those
of T. skrjabini, T. lacrymalis, and T. Callipaeda (Kikuchi
1976; Beelitz et al. 1997; Otranto et al. 2003a).
Two nipple-shaped phasmids were seen at the posterior
end of both sexes of T. rhodesi in this study. They are
involved in evaluation of the intensity of a given stimulus
859
and helping the worm to maintain itself in a suitable
environment (McLaren 1976).
Acknowledgements The author deeply thanks Mr. Samani and Mr.
Gerami, Faculty of Veterinary Medicine, Tehran University, Iran, who
were of enormous assistance in the collection of material examined in
this study. The author wishes to express her appreciation to Professor
Larry A. Arsenault, head of the electron microscope facility,
Department of Pathology and Molecular Medicine, McMaster Uni-
versity, Canada, for his assistance in providing all facilities during the
study. Thanks are due to Mr. Ernie Spitzer, chief technician, and all
the technicians in the electron microscope facility for their help. The
author is also grateful to Dr. Shigehiko Uni, Osaka City University,
Japan, for translating the Japanese article and helpful comments.
Finally, the author is especially obliged to Dr. Christine M. Budke,
Texas A&M University, USA, for her continuous support and critical
review of the manuscript.
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