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Labyrinthuloides haliotidis n.sp. (Protozoa: Labyrinthomorpha), a pathogenic

parasite of small juvenile abalone in a British Columbia mariculture facility

Article  in  Canadian Journal of Zoology · February 2011

DOI: 10.1139/z87-304

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 Labyrinthuloides haliotidis n.sp. (Protozoa: Labyrinthomorpha), a pathogenic parasite of
small juvenile abalone in a British Columbia mariculture facility
M. BOWER
SUSAN
Department of Fisheries and Oceans, Fisheries Research Branch, PaciJic Biological Station, Nanaimo, B.C., Canada
V9R 5K6
Received October 28, 1986

BOWER,S. M. 1987. Labyrinthuloides haliotidis n.sp. (Protozoa: Labyrinthomorpha), a pathogenic parasite of small juvenile
abalone in a British Columbia mariculture facility. Can. J. Zool. 65: 1996-2007.
Labyrinthuloides haliotidis n.sp. is an achlorophyllous eucaryotic protist that is pathogenic to juvenile abalone (Haliotis
kamtschatkana and Haliotis rufescens) less than 190 days of age (postsetting). Within the muscle and nervous tissue of the head
Can. J. Zool. Downloaded from www.nrcresearchpress.com by Depository Services Program on 06/06/13

and foot of susceptible abalone and in axenic nutrient culture media at 10°C, vegetative stages of L. haliotidis proliferated by
binary fission and produced ectoplasmic nets from sagenogenetosomes located on the cell periphery. When the abalone died and
the parasites were released from the decaying tissue or when culture forms were washed free of nutrient medium and placed in
sea water, internal multiple fission (sporulation) occurred within some cells, producing zoosporoblasts. After 24 to 72 h of
incubation at 10°C, the zoosporoblasts ruptured to release from 3 to about 10 infective biflagellated zoospores. After about 24 h of
active swimming, or on contact with a glass surface, the zoospores shed their flagella. Ultrastructure of vegetative stages and
zoospores related this species more closely to the thraustochytrids than to the labyrinthulids. Confusion still prevails concerning
the higher taxonomic affinities of this group of organisms. In keeping with recent publications on the taxonomy of the kingdom
Protozoa, L. haliotidis was considered to be a protozoan of the phylum Labyrinthomorpha and not allied with the fungi.

BOWER,S. M. 1987. Labyrinthuloides hliotidis n. sp. (Protozoa: Labyrinthomorpha), a pathogenic parasite of small juvenile
abalone in a British Columbia mariculture facility. Can. J. Zool. 65 : 1996-2007.
Labyrinthuloides haliotidis n.sp. est un protiste eucaryote sans chlorophylle parasite pathoghe des Haliotides (Haliotis
kamtschatkana et Haliotis rufescens) de moins de 190jours (aprks leur etablissement). Dans les tissus musculaires et nerveux de
la tCte et du pied des haliotides sensibles, de mCme que dans des milieux de culture axeniques a 10°C, les stades vCgCtatifs de L.
haliotidis peuvent prolifdrer par scission binaire et produisent des rdseaux ectoplasmiques issus des sagCnogCnCtosomes situCes
For personal use only.

en pkriphdrie de la cellule. Lorsque I'haliotide meurt et que les parasites sont IiberCs du tissu en decomposition ou lorsque les
formes cultivkes sont sorties du milieu de culture et placCes en eau de mer, il se produit des scissions internes multiples
(sporulation) a I'intCrieur de certaines cellules aboutissant a la formation de zoosporoblastes. Aprks 24 a 72 h d'incubation a
10°C, les zoosporoblastes se rompent et liberent de 3 a environ 10 zoospores infectieuses biflagellks. Aprks 24 h environ de nage
active ou en venant en contact avec une surface de verre, les zoospores rejettent leurs flagelles. D'aprks la microstructure des
stades vkgdtatifs et des zoospores, cette espkce se rapproche plus des Thraustochytrides que des Labyrinthulides, mais la
confusion rkgne encore au sujet des affinites taxonomiques plus genkrales de ce groupe d'organismes. Conformement aux
publications rCcentes sur la taxonomie du rkgne des Protozoaires, L. haliotidis est consider6 comme un protozoaire du phylum
des Labyrinthomorphes et ne s'apparente pas aux champignons.
[Traduit par la revue]

Introduction obtained from a mariculture facility on Vancouver Island. For

A new abalone mariculture facility on Vancouver Island,
British Columbia, has suffered high mortalities among small
juvenile abalone Haliotis kamtschatkana and Haliotis rufescens.
The foot muscle of moribund abalone was found to be heavily
infected with an invasive eucaryotic organism new to science
and allied with the thraustochytrids and labyrinthulids. The
taxonomy of this group of organisms is still in confusion (see
Discussion). Although they are commonly considered to be
fungi (Kumar 1979), I have decided to be consistent with recent
publications that placed these organisms in a separate phylum,
Labyrinthomorpha, within the kingdom Protozoa (Levine et al.
1980; Lauckner 1983; Pokorny 1985). Thus the descriptive
terminology used in this paper relates to protozoology and not
mycology.
The abalone parasite was cultured axenically and its life cycle
was completed in vitro and in vivo, with the formation of
infective biflagellated zoospores (Bower 1987a, 1987b). A
description of the new species, including characterization of all
life stages at the light microscope level and ultrastructurally, is
presented here. The new species is compared with other
thraustochytrids and labyrinthulids.
Materials and methods
Parasites in abalone
Small parasitized juvenile abalone (1 to 11 mm in shell length) were
examination, abalone were squashed in a drop of sea water between a
glass slide and cover slip and the compressed tissues were scanned with
a compound light microscope (250X magnification). Abalone larger
than 3 mm in shell length were removed from the shell before being
squashed.
To measure the vegetative stage of the parasite, a peripheral piece of
infected abalone tissue (unstained) was microscopically examined
under an oil immersion objective (1000x) and, with the aid of a
drawing tube, the circumference of 50 specimens was traced. The
maximum diameter to the nearest millimetre was measured for each
traced specimen and measurements were converted to micrometres
using a stage micrometer (10 pmldivision) which was also viewed
under oil immersion and traced. In addition, 50 late-stage zoosporo-
blasts on the edge of the decaying tissue as well as one clearly defined
zoospore within each zoosporoblast were drawn and measured.
For histological examination under the compound light microscope
(100 to 1 0 0 0 ~ )small
, infected abalone (approximately 2 mm in shell
length) were preserved in Bouin's fixative and embedded in paraffin
wax. Sections were cut at 7 pm and stained with Harris's modified
haematoxylin and 0.5% alcoholic eosin. For transmission electron
microscopic examination, intact, small, infected abalone (approxi-
mately 1.5 mm in shell length) were fixed in 3% formalin with 0.75%
glutaraldehyde in 0.1 M sodium cacodylate buffer at 4°C for 3 h, then
transferred to chilled sodium cacodylate buffer and warmed to room
temperature. Tissues were removed from the shells and the shells were
discarded. Within 5 days, the specimens were postfixed in 1% osmium
tetroxide in 0.1 M phosphate buffer, dehydrated in ethanol, and
embedded in Epon. Sections were stained with uranyl acetate followed
 BOWER: I
by lead citrate before being examined with a Philips electron
microscope 300.
Parasites in culture
The parasite was cultured aseptically at 10°C in medium designated
as MEM, which consisted of Eagle's minimum essential medium
containing Earle's salts, L-glutamine, 10% fetal calf serum, and
antibiotics consisting of 100 U penicillin/mL, 100 pg streptomycin/
rnL, and 0.25 pg FungizoneB/mL (GIBCO, Grand Island, NY 14072)
as described elsewhere (Bower 19876). Cultured parasites were
measured from living specimens and specimens fixed in 10% buffered
formalin using the same techniques described for the vegetative stages
in abalone. In addition to the single cells, each parasite of a pair
undergoing the late stages of binary fission was also measured
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individually.
Internal morphology was examined with a transmission electron
microscope (Philips electron microscope 300). For fixation, either the
cultured parasites were centrifuged (2000g) for 8 min and the pellet
resuspended in the same fixative and dehydration series used for the
abalone or the medium in a culture flask was replaced with a mixture of
2.5% glutaraldehyde and 1% osmium tetroxide in Millonig's phos-
phate buffer for 3 h of fixation at about 4°C followed by dehydration
with an ethanol series at room temperature. Specimens were embedded
in Epon, sectioned, and stained in a similar manner to the abalone
tissue.
To examine and photograph the ectoplasmic net produced in MEM
cultures, a drop of culture medium containing approximately five
parasites in every high power (400x) light microscope field was
aseptically placed between a slide and Vaseline-ringed cover slip. This
miniculture was incubated in a humidity chamber at 10°C for 4 days.
The resulting culture was photographed through the cover slip using a
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phase contrast oil immersion objective on a compound microscope.
The movement of the parasite along the plastic surface of MEM
cultures was monitored by placing a plastic culture flask in a small
circulating water bath at 10.5OC under a dissecting microscope (100 X )
equipped with a 35 mm camera. The flask was positioned such that
scratches in the plastic could be used to orient the specimens and
monitor their movement. Photographic exposures were taken at 15- to
60-min intervals over a 4.75 h period. The photographs were printed at
about 300x magnification and the movement of 10 parasites over the
time series was mapped and measured.
Zoospore production and examination
Zoospores were routinely produced by removing the vegetative cells
from MEM cultures and placing them in filtered sea water with
antibiotics as described elsewhere (Bower 1987b). The zoosporoblast
and zoospores were measured using the same techniques described for
the other culture stages except that before they were measured, these
stages were fixed in a final concentration of 7.5% formalin by adding
three volumes of 10% buffered formalin to one volume of parasites in
sea water. Details of external morphology of the zoospores were
examined with a scanning electron microscope (Etec Autoscan) by
placing several drops of formalin-fixed zoospores on a glass cover slip
and allowing the specimens to adhere to the glass by incubation in a
humidity chamber at 4OC for about 5 h. Cover slips were then
transferred, withoug being dried, through ethanol dehydration into
Genesolve@,critical-point dried in Freon 13, and sputter coated with
gold-palladium.
Internal ultrastructure was examined using the same procedure as for
the vegetative cells in abalone and MEM culture except zoosporoblasts
and zoospores were fixed by adding an equal volume of 6%
glutaraldehyde in Millonig's buffer (pH 7.3) to the parasites in
sea water. After 3 h of fixation, the parasites were washed 4 times with
Millonig's buffer by slow centrifugation (1500g for 8 min) to remove
the fine precipitate formed by the interaction between the fixative and
sea water. Some of these specimens were also negatively stained by
placing a drop of glutaraldehyde-fixed and washed zoospores on a
0.5% Formvar-coated grid and floating the grid on 0.5% phosphotunstic
acid for 30s. The grids were air dried and examined under the
transmission electron microscope to identify mastigonemes on the
flagellum.
Results
Labyrinthuloides haliotidis n. sp.
Description
Measurements in the following description were from 50
specimens found within or freed from dead and decaying
abalone except for the zoospores which were produced from
cultures.
Vegetative stages usually spheroidal in shape, diameter
7.2 + 0.8 (5.2-8.6) km (mean + standard deviation followed
by range in parentheses), rarely slightly ameboid; in nutrient
medium move slowly (average speed of 0.3 + 0.16 km/min for
10 individuals over 4.75 h); in abalone and nutrient medium,
divide by binary fission occasionally in quick succession to form
clusters; in sea water, binary fission retarded, some individuals
undergo synchronous multiple fission (sporulation) within the
sporangial wall to form zoosporoblasts (7.8 + 0.7 (6.2-
9.8) km); each zoospore develops within a membrane within
the sporangial wall and has a diameter of 4.3 + 0.5 (3.7-
7.1) km; uninucleate motile biflagellated zoospores slightly
oval, 4.7 + 0.5 (4.0-5.6) km long by 3.6 + 0.4 (2.8-4.4) km
wide, escape through rupture in sporangial wall; flagella
subapically attached, anterior flagellum 11.7 + 1.3 (9.4-
14.6) km with mastigonemes, posterior flagellum 6.7 + 0.9
(5.0-9.4) km with tapered tip and no mastigonemes; colonies
on 10% bovine serum agar medium greyish white, mainly
circular with an irregular leading edge.
HOSTS: Haliotis kamtschatkana and Haliotis rufescens juve-
niles up to at least 4.0 mm in shell length.
TRANSMISSION: Direct.
LOCALITY: Saanich Inlet, Vancouver Island, British Colum-
bia, Canada.
Type specimens are lodged in the National Museum of
Natural Science, Ottawa, Ontario, Canada, specimen Nos.
NMCP 1986-0893 to 0897. Specimen numbers correspond to
the following material: 0893, histological section of infected H.
kamtschatkana tissue preserved in Bouin's fixative, embedded
in paraffin wax, and stained with Harris's modified haema-
toxylin and 0.5% alcoholic eosin; 0894, histological section of
infected H. kamtschatkana tissue preserved in 3% formalin with
0.75% glutaraldehyde in sodium cacodylate buffer, postfixed in
1% osmium tetroxide in 0.1 M phosphate buffer, embedded in
Epon, and stained with methylene blue; 0895, vegetative stages
from aseptic cultures in minimum essential medium with 10%
fetal calf serum preserved in 10% buffered formalin; 0896,
zoosporoblasts, a few zoospores, and dying vegetative stages
from aseptic cultures placed in sterile sea water, preserved in
10% buffered formalin; 0897, zoospores, a few zoosporoblasts,
and dying vegetative stages from aseptic cultures placed in
sterile sea water, preserved in 10% buffered formalin.
Living cultures are lodged with Dr. Annemarie Ulken,
Alfred-Wegener-Institut fiir Polar und Meeresforschung, Post-
fach 120161, Columbusstrabe, D-2850 Bremerhaven, Federal
Republic of Germany, and Dr. Stephen T. Moss, Portsmouth
Polytechnic, School of Biological Sciences, King Henry I
Street, Portsmouth, Hampshire, England, U. K. PO 1 2DY.
Labyrinthuloides haliotidis in abalone
The vegetative forms of L. haliotidis occurred throughout the
tissues of the head and foot of the small juvenile abalone, but
were never observed in the visceral spiral. These vegetative
forms were usually forced from the weakened muscle tissue of
heavily infected abalone during the squashing procedure of
preparing fresh wet mounts. Under the light microscope,
specimens showed the characteristic light-refractile granules
 1998 CAN. J . ZOOL. VOL. 65, 1987
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FIG. 1. Wet mounted, unstained preparation of Labyrinthuloides haliotidis n.sp. liberated from an infected abalone, Haliotis kamtschatkana.
Dark, light-refractile granules (PG) and specimens undergoing binary fission (BF) are evident. Scale bar = 25 p.m. FIG. 2. Haematoxylin and eosin
stained histological section of juvenile H. kamtschutkam with L. haliotidis n.sp. (P) within the nerve ganglion and in surrounding tissues. Scale bar =
25 p.m. FIGS.3 and 4. Electron micrographs of the parasite L. haliotidis n.sp. within the muscle tissue of juvenile abalone (H. kamtschatkana).
Scale bars = 2.5 p.m. Fig. 3. Parasite (P), with golgi apparatus (GD) adjacent to the nucleus, lying next to nucleus (HN) of lysed host cell. Fig. 4.
Parasite with an ectoplasmic net (EN) and several mitochondria1 profiles (M) containing tubular cristae. A few nuclear pores (NP) are evident and a
golgi apparatus (GD) is located adjacent to the nucleus.

FIG. 5. Electron micrograph of a dividing Labyrinthuloides haliotidis n.sp. within the muscle tissue of a juvenile abalone, Haliotis
kamschatkana. A cell wall (CW) is evident between the two progeny and the parental cell wall (PCW) is disrupted on one side. Scale bar = 2.5 p.m.
FIGS.6-12. Electron micrographs of L. haliotidis n.sp. from liquid culture medium (minimum essential medium with 10% fetal calf serum). Fig.
6. Binary fission that occurred in rapid succession. Note that the parental cell wall (PCW) around both pairs has been disrupted in some places
whereas the wall around the pair on the left and that on the right is still intact. Also there are no electron-dense inclusions evident in the cytoplasm.
Scale bar = 2.5 p.m. Fig. 7 . Specimen from old culture with several electron-dense inclusions (EDI). A nuclear pore (NP) and mitochondrial
profiles (M) containing tubular cristae are evident. Scale bar = 2.5 p.m. Fig. 8. Sagenogenetosome (SA) on the periphery of a cell that was actively
producing ectoplasm nets (EN), which consist of a unit membrane with no evidence of cell organelles internally. Mitochondria1 profiles (M)
containing tubular cristae are evident adjacent to the sagenogenetosome. Scale bar = 1 p.m. Fig. 9. Golgi apparatus (GD) and centriole (C) adjacent
to the nucleus. Scale bar = 1 p.m. Fig. 10. A golgi apparatus (GD) that appears to be forming a cell wall scale (SC). Scale bar = 1 p.m. Fig. 11.
Structure resembling a lomasome (L). Scale bar = I p.m. Fig. 12. A specimen with a cell wall that appears to consist of several layers. Scale
bar = 1 p.m.
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1999
 2000 CAN. J. ZOOL. VOL. 65, 1987

TABLE1. Sizea (in Fm) of Labyrinthuloides haliotidis n.sp. from Haliotis kamtschatkana in vivo and in vitro at various
developmental stages

In vivo
In vitro
Freed from
Embedded in decaying abalone MEM nutrient Autoclaved or filtered
abalone tissue tissue medium sea water

Single vegetative cells

Dividing cells
Largest individuals
Smallest individuals
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Zoosporoblasts
Developing zoospores
within zoosporoblasts
Zoospores
Body length
Body width
Anterior flagellum
Posterior flagellum
"Except for zoospores in which specific measurements were taken, sizes are presented as the greatest diameter and recorded as mean + standard deviation with
range in parentheses. The sample size was 50 for each parameter except for specimens from MEM nutrient medium for which 300 individuals were measured for
each developmental stage.
For personal use only.

FIGS.13 and 14. Living unstained cultures of Labyrinthuloides
haliotidis n.sp. growing on a glass slide in a miniculture of minimum
essential medium with 10% fetal calf serum. Note the ectoplasmic nets
(EN) connecting the individuals of the colony and also a few cells with
an irregular border (IC). Swellings (SEN) were observed along the
ectoplasmic nets in older cultures. Fig. 13. Scale bar = 50 Fm. Fig. 14.
Scale bar = 25 Fm.
and were almost spherical in shape (Fig. 1). The only form of
division seen within intact abalone tissue was binary fission
(Fig. I). By the late stages of binary fission, when division was
almost complete, each individual of the pair was only slightly
smaller in size than nondividing individuals. When the abalone
died and began to decay, the vegetative stages of the parasite
were released. On entering sea water, a zoosporoblast was
formed by internal synchronous multiple fission (sporulation).
Mature zoosporoblasts were similar in size to the vegetative
stages (Table I). Using oil immersion light microscopy (1000x ),
the
bottom of a plastic culture flask, with five scratches, containing
minimum essential medium with 10% fetal calf serum. Time intervals
between consecutively numbered locations are 15,60, 31, 29, 38, 22,
30, 30, and 30 min, respectively.
each zoosporoblast appeared to contain from three to six but
usually four developing zoospores. The average maximum
diameter of individual zoospores within the zoosporoblast was
approximately half the diameter of the zoosporoblast (Table 1).
A few preparations of decaying infected abalone had a few
biflagellated zoospores swimming in the surrounding sea water.
On contact with the glass of the slide or cover slip, the zoospores
shed their flagella and rounded up from their swimming ovate
shape. The motile zoospores were similar in shape and size to
the zoospores produced by incubating MEM culture forms in
sea water.
Histologically, L . haliotidis vegetative forms were found
scattered throughout the nerves and muscles of the abalone foot
and head (Fig. 2) and occasionally in the gills. A clear space was
 BOWER: I 2001
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FIGS.16- 19. Electron micrographs of cultured Labyrinthuloides haliotidis n. sp. in sea water. Fig. 16. Dying and disintegrating forms. Scale
bar = 2.5 km. Fig. 17. A specimen undergoing synchronous multiple fission (sporulation) to form zoosporoblasts. Scale bar = 2.5 km. Fig. 18.
Zoosporoblast containing well-developed zoospores each within its own membrane wall, which is most evident around middle and two upper right
zoospores in the region of the cross section through the flagella. Scale bar = 2.5 km. Fig. 19. Mastigonemes (MA) extending from a flagellum of a
zoospore within the zoosporoblast. Note the mitochondrial profile (M) and ribosomes (R) within the cytoplasm of the zoospore. Scale bar = 1 km.

also noticed around most parasites (Fig. 2). The parasite was not
intracellular, but was capable of lysing its host's cells (Fig. 3).
The space around the parasite contained host cell debris (Fig. 4)
and scales peeling from the cell wall of the parasite (Figs. 3 , 4 ,
5). In abalone, the cytoplasm of L . haliotidis contained many
mitochondrial profiles with tubular cristae and vesicles with
various contents (Fig. 4). The nucleus had a few pores (Fig. 4)
and contained one distinct nucleolus (Figs. 3, 4, 5). A golgi
apparatus was usually located close to the nucleus (Figs. 3, 4).
During binary fission a cell wall was formed between the two
daughter cells, followed by the rupture of the surrounding
parent cell wall (Fig. 5).
Labyrinthuloides haliotidis in culture
Labyrinthuloides haliotidis grew well in MEM cultures at
10°C with a morphology similar to that in abalone. However,
the size of the organism in MEM cultures was more variable and
the mean greatest diameter (Table 1) was significantly larger
than the mean greatest diameter in abalone tissue (F-test, F =
155.7; df = 1,349; P < 0.05). There was no significant
difference in maximum diameter between fresh and formalin-
fixed material (F-test, F = 0.03; df = 1,198; P > 0.05).
Individuals of a pair in the late stages of binary fission were
usually slightly different in size (Table 1) with a significant
difference in the mean greatest diameter of the two progeny of
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FIGS.20-23. Electron micrographs of zoospores of Labyrinthuloides haliotidis n. sp. from sea water. Fig. 20. Scanning electron micrographs of
zoospores. Note the subapical attachment site of the two flagella, the coarse texture of the longer anterior flagellum where debris has attached to the
mastigonemes, and the thin tapered tip of the short posterior flagellum. Scale bar = 2.5 km. Fig. 2 1. Mastigonemes attached to about one-third of
the cross-sectioned flagellum circumference. Scale bar = 1 km. Fig. 22. Negatively stained whole mount illustrating the single thin terminal
extension (TTE) on the thicker basal portion of the mastigonemes. Scale bar = 1 km. Fig. 23. Section illustrating the small amount of
electron-dense substance in the kinetosome lumen (K) and the complex terminal plate - basal disc (BD). A vacuole close to the kinetosome appears
to be emptying (EX) and a mitochondrion (M) and scales (SC) sloughing from the cell wall are indicated. Scale bar = 1 km.

300 dividing pairs (Student's t-test, t = 7.9; df = 298; P <
0.001).
Ultrastructurally, the cytoplasm of cells in new, rapidly
growing cultures contained few vesicles (Fig. 6) but as the
culture aged and multiplication slowed down, the cytoplasm
became filled with electron-dense inclusions (Fig. 7). Sageno-
genetosomes were observed in some sections and, for specimens
fixed in situ, sections through extracellular ectoplasmic nets
were obtained (Fig. 8). These nets consisted of unit plasma
membranes with no evidence of cell organelles internally. As in
the abalone, the cytoplasm of L. haliotidis in vitro was filled
with rough endoplasmic reticulum and contained many mito-
chondria with tubular cristae. The nucleus had a distinct single
nucleolus and a few pores (Fig. 7). Golgi apparatus and
centrioles were occasionally observed adjacent to the nucleus
(Fig. 9). Electron-dense material was observed in the cisternae
of the golgi apparatus in one section (Fig. lo), which is
consistent with the supposed origin of wall scales in the
thraustochytrids (Darley et al. 1973; Perkins 1973; Porter 1974;
Perkins 1976; Chamberlain 1980; Moss 1980; McLean and
Porter 1982). Structures similar to lomasomes (Fig. 11) as
described by Perkins (1 969) were occasionally observed. There
was a variation in thickness of cell walls of different specimens
and in one case the cell wall appeared to consist of several layers
(Fig. 12). In MEM culture, L. haliotidis also divided only by
binary fission. However, divisions may occur in quick succes-
sion resulting in the formation of a few tetrads, but ultrastruc-
tural details of disintegrating cell walls reveal the time sequence
involved (Fig. 6).
Within 24 h of being placed in MEM culture, L. haliotidis
settled to the bottom of the flask and produced ectoplasmic nets
(Fig. 13). Swellings along the nets were observed in some
places in older cultures (Fig. 14). The parasites were motile and
moved independently (Fig. 15). In 4.75 h the greatest distance
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FIGS.24-26. Electron micrographs of zoospores of Labyrinthuloides haliotidis n.sp. from sea water. All scale bars = 1 Fm. Fig. 24. Section
through region of flagella attachment with the complex terminal plate - basal disc (BD) evident on both flagella. Fig. 25. Striated inclusion (SI)
adjacent to the kinetosome. Also note the ribosomes (R), the horseshoe-shaped mitochondrion (M) around the nucleus, and the scales (SC)
sloughing from the cell wall. Fig. 26. Section illustrating the kinetosome (K), the complex terminal plate - basal disc (BD), the horseshoe-shaped
mitochondrion (M) around the nucleus, and scales (SC) sloughing from the cell wall.
 2004 CAN. J. ZOOL. VOL. 6 5 , 1987
travelled by 1 (symbol x in Fig. 15) of 10 specimens was
196.3 pm. The fastest movement recorded was 1.13 pm/min
and the average speed of travel for the 10 individuals was
0.39 k 0.16 pm/min.
When MEM cultures of L. haliotidis were washed free of
culture medium and placed in flasks of sterile sea water, most of
the parasites reattached themselves to the bottom of the flask.
Some of the parasites subsequently died (Fig. 16). Others
underwent synchronous multiple fission (sporulation) to form a
zoosporoblast (Fig. 17). Within the walls of the zoosporoblast,
individual zoospores were formed, each within its own wall of
scales (Figs. 18, 19). The developing zoospore contained
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numerous vacuoles and one nucleus with a central nucleolus
(Fig. 18). The cytoplasm was filled with ribosomes but no rough
endoplasmic reticulum was observed and mastigonemes were
evident on the flagella (Fig. 19). Active mobile zoospores were
released when the cell wall of the zoosporoblast ruptured. Most
zoospores were released after 24-72 h of incubation in sterile
sea water at 10°C.
Zoospores were ovoid in shape, approximately 4.7 pm long
and 3.6 pm wide (Table 1) with two laterally attached flagella
(Fig. 20). Mastigonemes were found only on the longer anterior
flagellum and were attached to about one-third of the circum-
ference in cross section (Figs. 21, 25). Negative staining of
some mastigonemes showed a thin terminal extension on the
thicker basal portion (Fig. 22). The posterior flagellum was
shorter (about 6.7 pm in length, Table 1) and tapered at the tip
For personal use only.
(Fig. 20). The kinetosome-flagellum base contained a small
amount of electron-dense substance within the kinetosome
lumen (Figs. 23,26) and each flagellum had a complex terminal
plate - basal disc (Figs. 23, 24). The cytoplasm was filled with
ribosomes, and striated inclusions similar to those depicted by
Kazama (1972a) were found (Fig. 25). The only mitochondrion
observed in each zoospore was elongate and frequently found in
a horseshoe shape around the nucleus (Figs. 23,25,26). Scales
appeared to be sloughing from the thin cell wall of most
specimens (Figs. 23, 25, 26) and a few specimens appeared to
be voiding contents of vacuoles (Fig. 23).
Discussion
Labyrinthuloides haliotidis belongs to a group of achloro-
phyllous, eukaryotic protists known as thraustochytrids and
labyrinthulids. These organisms present formidable problems in
classification, particularly at the higher taxonomic levels
(Perkins 1976; Moss 1985). Perkins (1974a, 1976) indicated
that these organisms may have affinities with one or more of the
algal classes. Some workers (Goldstein 1973; Perkins 1976;
Gaertner 1977) have considered them to be lower fungi, while
others (Harrison and Jones 1974b; Alderman et al. 1974; Moss
1980, 1985), have placed them into two related orders,
Thraustochytriales and Labyrinthulales, of unspecified higher
taxonomic affinities. Olive (1975) proposed that these organ-
isms be grouped in the subphylum Labyrinthulina of the phylum
Gymnomyxa, kingdom Protista, with one class, Labyrinthulea,
one order, Labyrinthulida, and two familes, Labyrinthulidae
and Thraustochytriidae. More recently, Levine et al. (1980),
Lauckner (1983), and Pokorny (1985) included these organisms
in the subkingdom Protozoa and placed them in a separate
phylum, Labyrinthomorpha, with one class and one order as
proposed by Olive (1975).
In the original description, Perkins (1973) associated Laby-
rinthuloides with the only other genus, Labyrinthula, in the
group of labyrinthulids. This association was supported by
Harrison and Jones (1974b), but Alderman et al. (1974) and
Porter (1974) indicated that Labyrinthuloides has certain
features that appear to be intermediate between the Labyrin-
thulales and Thraustochytriales. Olive (1975) placed Labyrin-
thuloides in the family Thraustochytriidae leaving only one
genus, Labyrinthula, in the family Labyrinthulidae. Porter and
Kochert (1978) supported this proposal by showing that the
molecular weights of ribosomal RNA of two species of
Labyrinthuloides were similar to those of the thraustochytrids
and quite distinct from the labyrinthulids.
The morphology and behaviour of L. haliotidis, as observed
in the present study, supports the closer association of Labyrin-
thuloides with the thraustochytrids than with Labyrinthula. The
spherical shape of L. haliotidis is more like the round shape of
the majority of the thraustochytrids (thallus) than the elongate to
oval shape of Labyrinthula species (spindle cells). Perkins
(1976) claimed that the cell walls of thraustochytrids and
labyrinthulids are structurally similar, consisting of scales or
plates of electron-dense material. The cells of Labyrinthula
species are apparently surrounded by one or two overlapping
plates, although shadowed preparations of the plates have not
yet been obtained to confirm their existence (Perkins 1976). The
wall of the vegetative stage of L. haliotidis (Figs. 9, 12), on the
other hand, is similar to the cell wall in other Labyrinthuloides
species, Althornia crouchii (Perkins 1976), and Thraustochy-
trium sp. (Porter 1974). The multilamellate wall of the zoo-
sporoblast of L. haliotidis (Fig. 19) is similar to the walls of
zoosporoblasts (sporangium) and (or) vegetative stages (thalli)
of Thraustochytrium roseum illustrated by Chamberlain (1980),
Thraustochytrium striatum (Harrison and Jones 1974a),
Thraustochytrium kinnei (Harrison and Jones 1974b) , Japono-
chytrium sp. (Harrison and Jones 1974c), and Schizochytrium
aggregatum (Darley et al. 1973; Porter 1974). Labyrinthula
species apparently lack centrioles in the interphase vegetative
stage of the cycle, producing centrioles (protocentrioles) only
during zoosporulation (Perkins and Amon 1969; Porter 1972),
whereas centrioles were found in all stages of the life cycle of
Thraustochytriaceae as well as in Labyrinthuloides yorkensis
(Porter 1974; Perkins 1976). Centrioles were also observed in
nondividing vegetative forms of L. haliotidis (Fig. 9).
The morphology of the sagenogenetosome (=bothrosome,
Porter 1969) also differs between the two groups. In Labyrin-
thula species, the sagenogenetosome is located at the base of a
pit-like plasmalemmal invagination whereas in the thrausto-
chytrids and in Labyrinthuloides yorkensis, the sagenogeneto-
some is flush with the surface of the cell (Harrison and Jones
1974c; Olive 1975; Moss 1980). The sagenogenetosome of L.
haliotidis is also flush with the surface of the cell (Fig. 8) with
ultrastructural details similar to those of Labyrinthuloides
minuta as illustrated by Perkins (1972). Porter (1972) also
indicated that sagenogenetosomes occur between dividing
Labyrinthula where they were involved in the division process.
No workers have mentioned the presence of sagenogeneto-
somes between dividing pairs of Labyrinthuloides species or
thraustochytrids and none was observed in the present study on
L. haliotidis.
The sagenogenetosomes are involved in the production of the
ectoplasmic net. In Labyrinthula species the ectoplasmic net
("slimeway" or extracellular matrix membrane) envelopes or
enrobes the colony of individuals (Hohl 1966; Porter 1969,
1972) whereas individuals of Labyrinthuloides species and
thraustochytrids each produce a separate ectoplasmic net and
the net does not enrobe the individual. The latter condition was
 BOWER: I
also exhibited by L. haliotidis. Apparently, the ectoplasmic net
is used in the motility of Labyrinthula and Labyrinthuloides
species and this common feature has been used to group these
two genera together (Perkins 1976; Moss 1980). However,
Labyrinthula species can move singly or as a group through the
ectoplasmic net (Olive 1975) whereas individual L. haliotidis
and probably other Labyrinthuloides species usually move
independently and not in concert with other individuals (Fig.
15) and apparently glide on top of the ectoplasmic net although
the mechanism of movement is not fully understood. The rate of
movement is also different. Labyrinthula species in some cases
move as rapidly as 200 km/min (Olive 1975). The maximum
Can. J. Zool. Downloaded from www.nrcresearchpress.com by Depository Services Program on 06/06/13
speed recorded in this study for L. haliotids was about
1 km/min and the fastest movement for a Labyrinthuloides
species was reported by Watson and Raper (1957) as 4 to
5 km/min for L. (= Labyrinthula) minuta. Although thrausto-
chytrids usually are not motile, under certain culture conditions
young cells of species of the genera Thraustochytrium and
Schizochytrium may show limited movement by means of the
ectoplasmic net (Kazama et al. 1975; Olive 1975).
Although taxonomic significance has been attached to zoo-
spore morphpology in fungal phylogeny, and zoospores of
many species of thraustochytrids and labyrinthulids have been
examined, before this study, Labyrinthuloides zoospores had
not been described. Zoospores of L. haliotidis are similar in
length and width to those of three species (Labyrinthula
algeriensis, Thraustochytrium sp., and Schizochytrium aggre-
For personal use only.
gatum) listed by Perkins (1976), but the anterior and posterior
flagella are, on average, slightly shorter. The posterior flagellum
of L. haliotidis, like that of thraustochytrids and labyrinthulids,
is shorter than the anterior flagellum and the posterior flagellum
has a tapered tip as was noted for Labyrinthula sp. (Amon and
Perkins 1968), Thraustochytrium sp. and Althornia crouchii
(Kazama 1974), and Schizochytrium aggregatum (Perkins
1 9 7 4 ~ ) Also,
. as in thraustochytrids and labyrinthulids, L.
haloitidis has mastigonemes only on the anterior flagellum.
However, the location of the mastigonemes on the flagella of
zoospores in both groups has been described as being in two
rows (Amon and Perkins 1968; Perkins 1974a, 1976; Porter
1974). The negatively stained zoospores of L. haliotidis (Fig.
22) may also suggest a similar configuration of mastigonemes,
but cross sections of the anterior flagellum (Figs. 21, 25)
illustrate that the mastigonemes originate on a band that covers
about one-third of the circumference of the flagellum. Kazama
(1974) suggested that mastigonemes of both thraustochytrids
and labyrinthulids have fine hair-like extensions on the distal
tip. In Thraustochytrium sp. the tip of each mastigoneme has
two hairs whereas the mastigonemes of Labyrinthula sp.
(Kazama 1974) and L. haliotidis (Fig. 22) have only one
hair-like extension. Another possible difference between thrau-
stochytrids and L. haliotidis lies in the loss of the flagella by the
zoospores. Goldstein (1963) and Kazama et al. (1975) described
the zoospores of Thraustochytrium as "retracting" the out-
stretched flagella on settlement for development of the vegetative
stage. In contrast, at settlement, zoospores of L. haliotidis
quickly shed both their flagella, presumably by severing them at
the basal plate. This feature should be examined in other species
of thraustochytrids and labyrin thulids.
Zoospores of L. haliotidis contained striated inclusions (Fig.
25) morphologically similar to those illustrated by Kazama
(1972~)in a Thraustochytrium species that resembles T. moti-
vum. However, unlike the striated inclusions of T. motivum,
which seemed to be associated with a sagenogenetosome-like
2005
(bothrosome-like) organelle, the striated inclusions of L. halio-
tidis appeared to be associated with the kinetosome and thus
reminiscent of the striated fiber apparatus of Labyrinthula sp.
illustrated by Perkins and Amon (1969).
The kinetosome complex of L. haliotidis zoospores is unique
in that the lumen of the kinetosome complex contains only a
small amount of electron-dense substance (Figs. 23,26) and not
a cyclindrical inclusion or electron-opaque granule like those
reported for Thraustochytrium sp. (Kazama 1972b) and Laby-
rinthula sp. (Perkins and Amon 1969). In L. haliotidis the
kinetosome complex consists of a thick basal plate or disc at the
proximal end of the flagellum which may include the cyclind-
rical inclusion abutting the terminal plate (Figs. 23, 24, 25).
Vacuoles that appear to be excreting contents (Fig. 23) have
not been observed in other zoospores. Harrison and Jones
(1974b) suggested that the presence of membranous vesicles in
the zoospores of Thraustochytrium kinnei, Thraustochytrium
sp., Althornia crouchii, and Labyrinthuloides yorkensis could
have resulted from complete exhaustion of electron-dense lipid
material in earlier developmental stages. This may be the case
with L. haliotidis, which also appear to be capable of voiding
the undigested remnants by excretion.
Two features can be used to separate L. haliotidis zoospores
from those of Labyrinthula species and to align them with
zoospores of the thraustochytrids. First, like the zoospores of
Thraustochytrium sp. illustrated by Kazama (1973, 1974) and
Kazama et al. (1975), and zoospores of Thraustochytrium sp.,
Schizochytrium aggregatum, and Althornia crouchii illustrated
by Perkins (1976), the cell wall of L. haliotidis zoospores
consists of sloughing scales (Figs. 23-26). Scales on cell walls
of zoospores of Labyrinthula species have not yet been
described, nor are they indicated in the figure presented by
Perkins (1976). The second differentiating feature is the
absence in L. haliotidis zoospores of an eyespot (stigma) and
flagellar swelling on the posterior flagellum adjacent to the
eyespot granule, as observed in Labyrinthula species (Perkins
and Amon 1969; Porter 1974; Perkins 1976). Zoospores of
thraustochytrids also lack the eyespot and flagellar swelling
(Porter 1974).
Labyrinthuloides haliotidis is readily differentiated from the
species of the other six genera of thraustochytrids. From
Thraustochytrium, consisting of about 13 species, it differs in
being motile throughout its vegetative life and in certain
characteristics of the sagenogenetosomes and zoosporoblast. In
Thraustochytrium species, the ectoplasmic net originates from a
cluster of many sagenogenetosomes to form a single trunk
(Moss 1980). Also, a basal rudiment remains in some zoosporo-
blasts of each species after zoospore discharge, and this basal
rudiment can grow to form a new zoosporoblast with a new
basal rudiment (Sparrow 1968; Gaertner 1972; Kazama et al.
1975). In L. haliotidis, single sagenogenetosomes appear to be
randomly spread over the surface of the cell, and a basal
rudiment has not been observed. Ulkenia, consisting of six
species, is characterized by the naked stage of the protoplast and
ameboid migration before cleavage and formation of zoospores
(Gaertner 1977; Moss 1980). In L. haliotidis, a naked protoplast
was not observed and the vegetative stage did not appear to have
ameboid migration before zoosporoblast formation. The re-
maining four genera of thraustochytrids are all monotypic.
Althornia crouchii is free floating and lacks an ectoplasmic net,
and Aplanochytrium kerguelensis reproduces only by the
formation of aplanospores or nonflagellated zoospores (Moss
1980). Japonochytrium marinum has a dilation, the apophysis,
 2006 CAN. J. ZOOL. VOL. 65. 1987
on the ectoplasmic net at the base of the vegetative stage
(thallus), and the wall of the zoosporoblast (sporangium) is
persistent after the zoospores have been liberated through an
apical pore (Gaertner 1972; Moss 1980). Apophyses were not
observed at the base of the ectoplasmic nets of L. haliotidis.
However, in this species, swellings were noticed along the
length of the ectoplasmic net (Fig. 14) but they may have
resulted from ectoplasmic net breakdown because they were not
observed in young expanding colonies (Fig. 13). Also, the
zoospores of L. haliotidis did not seem to be liberated from an
apical pore but rather exited from a tear at any location on the
zoosporoblast wall. Although it was difficult to observe, the thin
Can. J. Zool. Downloaded from www.nrcresearchpress.com by Depository Services Program on 06/06/13
zoosporoblast wall may have persisted for a short time after all
the zoospores escaped, but it most likely disintegrated shortly
thereafter because intact cell walls were never observed in
seawater cultures that contained over 10' zoospores/mL. The
final genus of thaustochytrids, Schizochytrium, is characterized
by formation of tetrads in the vegetative stage, a result of
successive division of zoospores. Vegetative stages described
as an amorphous mass ranging between 30 and 140 pm diameter
that eventually separate into zoospore-producing segments have
also been noted (Goldstein and Belsky 1964; Moss 1980).
Discrete tetrads of vegetative stages were rarely observed in L.
haliotidis. In uncrowded conditions it appeared as though
divided pairs migrated away from each other soon after division
was completed. Also, amorphous masses were never observed
in abalone or MEM cultures, although L. haliotidis tended to
For personal use only.
form mounds on agar plates not flooded with sea water (Bower
1987b).
Labyrinthuloides haliotidis satisfies the generic description
of Labyrinthuloides given by Perkins (1973) except that
multiple fission in vegetative stages always results in the
production of a zoosporoblast (sporangium) that produces
biflagellated zoospores and not plasmodia. Amoeboid stages
were not observed; however, their presence is not an essential
feature of the genus. Another feature proposed as being
characteristic of the genus, but not mentioned in the official
generic description, is the presence of a membrane-inclusion
array located within a vesicle in the cytoplasm of the vegetative
stage (Perkins 1973). Membrane-inclusion arrays like those
illustrated by Perkins (1973) were not observed in the vegetative
stage of L. haliotidis; however, morphologically similar striated
inclusions were observed in the zoospores (Fig. 25).
To date, Labyrinthuloides has consisted of four species: L.
yorkensis was originally obtained from the mantle cavity of an
oyster (Crassostrea virginica) but was also isolated from water
samples, sediments, sand, and detritus in the estuary of the
York River, Virginia (Perkins 1973). Labyrinthuloides minuta
(Perkins 1974b), originally described as Labyrinthula minuta
by Watson and Raper (1957), was isolated from Ulva lactuca
and other algae collected from the Atlantic Ocean in the region
of Woods Hole, Massachusetts. Labyrinthuloides (= Labyrin-
thula) saliens, isolated from the seagrass Halophila engle-
mannii (Quick 1974a), and Labyrinthuloides schizochytrops,
isolated from the seagrass Halodule wrightii (Quick 1974b),
were both collected near Florida.
The nature of the symbiotic relationship between these
Labyrinthuloides species and their plant hosts was not men-
tioned for L. minuta. It was thought to be nonpathogenic and
nonparasitic for L. saliens (Quick 1974a), and remains open to
question for L . schizochytrops (Quick 1974b) . Although L .
yorkensis was recovered from the mantle cavity of oysters and
grew in cultures on oyster (C. virginica) and crab (Panopeus
herbstii) tissues, it was not thought to be pathogenic to
invertebrates (Perkins 1973). Thus, L. haliotidis is the first
species of this genus to be described as an invertebrate parasite
that is lethal to small juvenile abalone (Bower 1 9 8 7 ~ ) .
The morphology of L. haliotidis is slightly different from that
of the other four species in the genus. Labyrinthuloides
haliotidis is spherical and on the average, according to
measurements given by Quick (1974b), seems to be slightly
larger than the sperical L. schizochytrops (4-7 pm) and the
spherical to oval L. yorkensis (3-6 pm) but similar in size to
the ovoid species L. minuta (6- 10 pm) and L. saliens (5-8 pm).
The zoosporoblasts of L. haliotidis are smaller than the
sporangium (probably an equivalent life cycle stage of the
zoosporoblast although zoospores were not observed for two
Labyrinthuloides species) of the other four species as reported
by Quick (1974b). Also, plasmodia have been described for the
other four species but were not observed in L. haliotidis
although a few spherical cells with an indistinct and somewhat
irregular border were seen in MEM cultures (Fig. 14). The
viability of these cells of L. haliotidis was not determined.
Multiplication of L. yorkensis can occur by successive
division or progressive cleavage to produce between 2 and 64
daughter cells (Perkins 1973). Vegetative stages of L. schizo-
chytrops, L. minuta, and L. saliens each produce four, two to
four, and two to eight daughter cells, respectively, on division
of one parent cell (Perkins 1974b; Quick 1974a, 1974b). In the
present study, L. haliotidis only divided by binary fission in
abalone or nutrient medium, although this division could
occasionally occur in quick succession to form tetrads (Fig. 6).
Unlike the other four species that showed alternations of culture
forms between vegetative growth (spindle cell colonies) and
zoosporoblast production (sporangial colonies), L. haliotidis
only produced zoosporoblasts, with multiple internal fission, in
sea water when presumably little or no nutrients were available
for vegetative growth. These zoosporoblasts usually appear to
contain from 3 to 6 zoospores under the light microscope
(1000 x ), although as many as 10 zoospores were seen using the
electron microscope (Fig. 18). Mother cells containing at least
64 cells in L. yorkensis (Perkins 1973) and over 50 cells in
L. saliens and L. schizochytrops (Quick 1974a, 1974b) have
been described.
Experimental studies (Bower 1987a) have demonstated that
almost all juvenile abalone (up to 4.0 mm in shell length and less
than 190 days old) exposed to L. haliotidis, isolated from
infected abalone and from axenic cultures, succumb to the
infection within 10 days, with numerous vegetative stages of the
parasite throughout the head and foot tissues. Apart from reports
of undescribed thraustochytrid and labyrinthula-like organisms
observed on lesser octopus Eledone cirrhosa (Polglase 1980),
dendronotacean nudibranchs Tritonia diomedea (Mclean and
Porter 1982), and short-finned squid Illex illecebrosus (Jones
and O'Dor 1983), L. haliotidis is the first described thrausto-
chytrid that is a pathogenic parasite of a marine invertebrate.
Further experimental work is required to identify the host
specificity and geographic distribution of this pathogen.
Acknowledgments
I wish to thank the following persons for their contributions
during this study: Johanne LalibertC and Sally Goldes for their
technical assistance in the maintenance of the abalone and
parasite cultures; Dr. C. L. Singla, at the University of Victoria,
for his assistance in the electron microscopic examination of the
parasite; and Guy Whyte, of Pacific Trident Mariculture Ltd.,
 BOWER: I
who supplied the parasitized abalone. Drs. L. Margolis, J.
Polglase, F. Perkins, and L. Ellis provided valuable comments
o n earlier drafts.
ALDERMAN, D. J., HARRISON, J. L., BREMER,G. B., and JONES,
E. B. G. 1974. Taxonomic revisions in the marine biflagellate fungi:
the ultrastructural evidence. Mar. Biol. (Berlin), 25: 345-357.
AMON,J. P., and PERKINS, F. 0 . 1968. Structure of Labyrinthula sp.
zoospores. J . Protozool . 15: 543-546.
BOWER,S. M. 1987a. Pathogenicity and host specificity of Labyrin-
thuloides haliotidis (Protozoa: Labyrinthomorpha), a parasite of
juvenile abalone. Can. J. Zool. 65. This issue.
Can. J. Zool. Downloaded from www.nrcresearchpress.com by Depository Services Program on 06/06/13
1987b. Artificial culture of Labyrinthuloides haliotidis (Pro-
tozoa: Labyrinthomorpha), a pathogenic parasite of abalone. Can. J.
Zool. 65. This issue.
CHAMBERLAIN, A. H. L. 1980. Cytochemical and ultrastructural
studies on the cell walls of Thraustochytrium spp. Bot. Mar. 23:
669-677.
DARLEY,W. M., PORTER,D., and FULLER,M. S. 1973. Cell wall
composition and synthesis via golgi-directed scale formation in the
marine eucaryote, Schizochytrium aggregatum with a note on
Thraustochytrium sp. Arch. Mikrobiol. 90: 89- 106.
GAERTNER, A. 1972. Characters used in the classification of thrausto-
chytriaceous fungi. Veroeff. Inst. Meeresforsch. Bremerhaven, 13:
183-194.
1977. Revision of the Thraustochytriaceae (lower marine
fungi). I. Ulkenia nov. gen., with description of three new species.
Veroeff. Inst. Meeresforsch. Bremerhaven, 16: 139-157.
GOLDSTEIN, S. 1963. Development and nutrition of new species of
For personal use only.
Thraustochytrium. Am. J. Bot . 50: 27 1-279.
1973. Zoosporic marine fungi (Thraustochytriaceae and
Dermocystidiaceae). Annu. Rev. Microbiol. 27: 13-26.
GOLDSTEIN, S . , and BELSKY,M. 1964. Axenic culture studies of a new
marine phycomycete possessing an unusual type of asexual repro-
duction. Am. J. Bot. 51: 72-78.
HARRISON, J. L., and JONES,E. B. G. 1974a. Zoospore discharge in
Thraustochytrium striatum. Trans. Br. Mycol. Soc. 62: 283-288.
1974b. Ultrastructural observations on the formation of
zoospores in Thraustochytrium kinnei Gaertner. Trans. Mycol. Soc .
Jpn. 15: 273-288.
1974c. Ultrastructural aspects of the marine fungus Japono-
chytrium sp. Arch. Microbiol. 96: 305-3 17.
HOHL,H. R. 1966. The fine structure of the slimeways in Labyrin-
thula. J. Protozool. 13: 4 1-43.
JONES,G. M., and O'DOR, R. K. 1983. llltrastructural observations on
a thraustochytrid fungus parasite in the gills of squid (Illex
illecebrosus Lesueur). J. Parasitol. 69: 903-9 1 1 .
KAZAMA,F. Y. 1972a. Ultrastructure of Thraustochytrium sp.
zoospores. 11. Striated inclusions. J. Ultrastruct. Res. 41: 60-66.
1972b. Ultrastructure of Thraustochytrium sp. zoospores. I.
Kinetosome. Arch. Mikrobiol. 83: 179- 188.
1973. Ultrastructure of Thraustochytrium sp. zoospores. 111.
Cytolysomes and acid phosphatase distribution. Arch. Mikrobiol.
89: 95-104.
1974. Ultrastructure of Thraustochytrium sp. zoospores. IV.
External morphology with notes on the zoospores of Schizochytrium
sp. Mycologia, 66: 272-280.
KAZAMA, F. Y., ZACHARY, A. L., and SCHORNSTEIN, K. L. 1975.
Observations on Thraustochytrium sp.: development and behavior
in culture. Can. J. Bot. 53: 360-374.
KUMAR,S. R. 1979. Observations of the life cycle and movement of
the thraustochytrid Ulkenia amoeboidea (Bahnweg and Sparrow)
from the North Sea. J. Protozool. 26: 564-566.
LAUCKNER, G. 1983. Diseases of Mollusca: Bivalvia. In Diseases of
marine animals. Vol. 11. Introduction, Bivalvia to Scaphopoda.
View publication stats
2007
Edited by 0 . Kinne. Biologische Anstalt Helgoland, Hamburg,
Federal Republic of Germany. pp. 477-961.
LEVINE,N. D., CORLISS,J. O . , COX,F. E. G., DEROUX,G., GRAIN,
J., HONIGBERG, B. M., LEEDALE,G. F., LOEBLICH,A. R., 111,
LOM,J., LYNN,D., MERINFELD, E. G . , PAGE,F. C . , POLJANSKY,
G., SPAGUE,V., VARVA,J., and WALLACE, F. G . 1980. A newly
revised classification of the Protozoa. J. Protozool. 27: 37-58.
MCLEAN,N., and PORTER,D. 1982. The yellow-spot disease of
Tritonia diomedea Bregh, 1894 (Mollusca: Gastropoda: Nudibran-
chia): encapsulation of the thraustochytriaceous parasite by host
amoebocytes. J. Parasitol. 68: 243-252.
Moss, S. T. 1980. Ultrastructure of the endomembrane-sagenogeneto-
some-ectoplasmic net complex in Ulkenia visurgensis (Thrausto-
chytriales). Bot. Mar. 23: 73-94.
1985. An ultrastructural study of taxonomically significant
characters of the Thraustochytriales and the Labyrinthulales. Bot. J.
Linn. Soc. 91: 329-357.
OLIVE,L. S. 1975. The mycetozoans. Chapt. 7 . Labyrinthulina
(labyrinthulas and thraustochytrids). Academic Press, New York.
pp. 215-281.
PERKINS,F. 0 . 1969. Ultrastructure of vegetative stages in Labyrin-
thomyxa marina (= Dermocystidium marinum), a commercially
significant oyster pathogen. J. Invertebr. Pathol. 13: 199-222.
1972. The ultrastructure of holdfasts, "rhizoids", and "slime
tracts" in thraustochytriaceous fungi and Labyrinthula spp. Arch.
Mikrobiol. 84: 95- 1 18.
1973. A new species of marine labyrinthulid Labyrinthuloides
yorkensis gen. nov. spec. nov. -cytology and fine structure. Arch.
Mikrobiol. 90: 1 - 17.
1974a. Phytogenetic considerations of the problematic
thraustochytriaceous-labyrinthulid-Dermoqstidium complex based
on observations of fine structure. Veroeff. Inst. Meeresforsch.
Bremerhaven Suppl. 5: 45-63.
1974 b . Reassignments of Labyrinthula minuta to the genus
Labyrinthuloides. Mycologia, 66: 697-702.
1976. Fine structure of lower marine and estuarine fungi. In
Recent advances in aquatic mycology. Edited by E. B. G. Jones.
Paul Elek Ltd., London. pp. 279-3 12.
PERKINS,F. O., and AMON,J. P. 1969. Zoosporulation in Labyrin-
thula sp. ; an electron microscope study. J. Protozool. 16: 235-257.
POKORNY, K. S . 1985. Phylum Labyrinthomorpha. In An illustrated
guide to the Protozoa. Edited by J. J. Lee, S. H. Hutner, and E. C.
Bovee. Society of Protozoologists, Lawrence, Kansas. pp. 318-321.
POLGLASE, J. L. 1980. A preliminary report on the thraustochytrid(s)
and labyrinthulid(s) associated with a pathological condition in the
lesser octopus Eledone cirrhosa. Bot. Mar. 23: 699-706.
PORTER,D. 1969. Ultrastructure of Labyrinthula. Protoplasma, 67:
1-19.
1972. Cell division in the marine slime mold, Labyrinthula
sp., and the role of the bothrosome in extracellular membrane
production. Protoplasma, 74: 427-448.
1974. Phylogenetic considerations of the Thraustochytriaceae
and Labyrinthulaceae. Veroeff. Inst. Meeresforsch. Bremerhaven
Suppl. 5: 19-44.
PORTER,D., and KOCHERT,G . 1978. Ribosomal RNA molecular
weights and the phylogeny of Labyrinthula. Exp. Mycol. 2:
346-35 1 .
QUICK,J. A. 1974a. A new marine Labyrinthula with unusual
locomotion. Trans. Am. Microsc. Soc. 93: 52-61.
1974 b . Labyrinthuloides schizochytrops n. sp., a new marine
labyrinthula with spheroid "spindle" cells. Trans. Am. Microsc.
SOC.93: 344-365.
SPARROW,VON F. K. 1968. Remarks on the Thraustochytriaceae.
Veroeff. Inst. Meeresforsch. Bremerhaven, 3: 7-18.
WATSON,S. W., and RAPER,K. B. 1957. Labyrinthula minuta sp.
nov. J. Gen. Microbiol. 17: 368-377.