JKAU:
J. KAU:Eng.
Trichodiniasis
Mar.
Sci.,
Sci.,
vol.
vol.
11in
10,
no.
Farmed
pp.
1, pp.
157-168
Freshwater
3-17(1419
(1419A.H.
Tilapia...
A.H.//1999
1999A.D.)
A.D.)
157
Trichodiniasis in Farmed Freshwater Tilapia in
Eastern Saudi Arabia
MOHAMED ABD EL-AZEZ HASSAN HASSAN
Fisheries Research Center, Al-Qateef, Saudi Arabia
ABSTRACT. The present study is aimed at screening for the presence of Trichodina sp. among Tilapia spp. (Oreochromis niloticus and Oreochromis aureus)
collected from various fish farms in the eastern Province of Saudi Arabia during January, 1995-May, 1997. Out of 675 fishes collected only 335, (49.6%)
were infested with Trichodina. The highest prevalence was observed in the
spring and winter followed by autumn and summer. In general, the intensity of
infection was low except in cases where outbreaks of Trichodiniasis endangered the survival of fish in some ponds. In light infestation Trichodina is usually present on gills, fins and skin of apparently healthy fish. Clinical signs of
Trichodiniasis only appear on fish with heavy infections and cases of moderate
ones that are usually exposed to one or more stress factors including, rough
handling during transportation from ponds, overcrowdness, malnutrition, high
of free ammonia and low of oxygen concentration. Clinical signs of Trichodiniasis in fish are sluggish movement, loss of appetite, black colouration, necrosis
and ulcer on different parts of the body, detached scales and excessive accumulation of mucous in gill pouches. The most obvious histopathological changes
in diseased fish were sloughing of the epidermal layer, aggregation of leucocytes and melanine-carrying cells (between the dermis and hypodermis) and proliferative changes including hyperplasia and hypertrophy of the epithelial lining cells of gill filaments which resulted in fusion of secondary lamellae.
Control of Trichodiniasis, has been achieved by formalin bath treatment at a
concentration of 250 ppm for one hour.
Introduction
The concept of aquaculture development as a potential source of food supply is receiving much attention in the Kingdom of Saudi Arabia. This has resulted in rapid growth in
fish farming practice.
The intensification of such fish culturing creates disease problems that originate from
overcrowdness (Sarig, 1968a) or deteriorating water quality such as unsuitable water
temperature (Dujin, 1973), pH (Kugel et al., 1990), carbon dioxide and free ammonia
concentrations.
157
158
Mohamed Abd El-Azez Hassan Hassan
It is usually known that external parasites constitute the largest group of pathogenic
organisms in warmwater fish (Snieszko and Axelrod, 1971). The majority of fish ectoparasitic protozoa are commensals but some of them may produce serious diseases and
mortality especially in fry and fingerlings. The toxonomy and pathological effects of
these parasites have been dealt with by Lom and Hoffman (1964), Wellborn (1967) and
Hoffman (1978), Amlacher (1970), Molnar (1971), Kennedy (1974) and Van As and
Linda Basson (1983).
Parasitic protozoa have a direct life cycle that enables them to spread rapidly and
widely in their hosts (Kudo, 1966). Such rapid growth and multiplication of these parasites greatly increase their rate of infestation among fish causing mass mortality.
Trichodina are a very common ectoparasites which in most cases are pathogenic to
both freshwater and marine fish (Wellborn, 1967). Dogiel (1961) reported that trichodiniasis is caused by Trichodina spp.; the infection being stimulated by the high density
of fish in ponds. Under such conditions, it may be an epizootic. In cases of stress, several species of trichodina may become pathogenic interfering with feeding and respiration
of small fish (Sarig, 1968b and Ahmed, 1976).
The lesions mostly induced by this parasite are hyperplasia and necrosis of the epidermal cells. These cells are usually subjected to severe attacks by parasites. This results in cellular growth and excess mucus production (Ahmed, 1976). Many authors
have observed a complete destruction of gill epithelium in similar cases (Paperna, 1980;
McArdle, 1984; and Eisa et al., 1985).
For the control of Trichodiniasis, formalin is recommended by many authors that
have experimented with different concentrations ranging between 166 to 250 ppm applied directly in short-term bath (Amlacher, 1970; Klinke, 1973; Paperna, 1980; and
McArdle, 1984).
The present work was aimed to achieve the following objectives: First, to investigate
the incidence of Trichodina in freshwater Tilapia (Oreochromis niloticus and Oreochromis aureus) in different farms in the eastern Province of Saudi Arabia. Secondly, to examine and evaluate the magnitude of injury that induced by this parasite in infected tissues and organs of the fish. Thirdly, to test efficacy of formalin in controlling the
parasite.
Material and Methods
In the present investigation, about 675 tilapias were collected regularly from various
ponds of fry, fingerlings, growing and brood stock, during a fish health survey of different fish farms in the eastern Province of Saudi Arabia and also during emergency cases
where some fish mortalities were observed during the period of January, 1995-May,
1997.
After observation of fish behaviour in various ponds, collected samples were transported (alive) to the laboratory of Fisheries Research Center, Al-Qateef, in large plastic
bags which were aerated upon arrived to the laboratory, fish were maintained in aerated
Trichodiniasis in Farmed Freshwater Tilapia...
159
aquaria. The fish were checked daily for clinical signs, according to the method described by Amlacher (1970).
Skin fins and gills smears of these fish were made and carefully searched for Trichodina sp. The severity of infection with Trichodina sp. was estimated on an arbitrary
scale using ordinary microscope under a 10 × objective lens where no parasites were observed ( – ), low infection with single parasite in several fields of vision (+), medium infection with 2 to 5 parasites per field of vision (++) and severe infection with more parasites per field of vision (+++). After examining parasites alive, they were fixed to
microscope slides with methyl alcohol then stained with Giemsa according to Lucky
(1977).
Gills and skin specimens from clinically infested fish were fixed in phosphate buffered formalin, trimmed and prepared for histopathological examination. Specimens
were embedded in paraffin wax and sectioned using a rotary microtome at 5-7 µm thickness. Specimens were stained with haematoxylin and eosin (Hibiya, 1982).
During outbreaks of Trichodiniasis among fish in some ponds, dissolved oxygen concentrations were measured with Orion model 850 oxygen meter and ammonia (as ammonia-nitrogen) was determined by the Nessler method (Hach model DR/2000).
Clinically infested fish in ponds were treated by addition of formalin at a concentration of 250 ppm for one hour (Klinke and Elkan, 1965). Treated fish were checked after
24 hours.
Results
Identification of the parasite
Trichodina were recorded from the body surface, fins and gills of Tilapia. In fresh unstained preparation, the parasite is very motile and appears as a circular or bell-shaped
ciliated organism. The adhesive disc (saucer-shaped), several circular rows of cilia and
a circle of more centrally lying hooklets could be easily seen in preparations stained
with Giem’s stain, Fig. 1.
Epizootology and clinical picture of the infection
Out of 675 fish, 335 were found infested with Trichodina (49.6% infestation). Data
indicate that the highest incidence of infestation was found in growing fish (61.11%)
followed by fingerling (54.28%), Brood stock (46.87%) and lastly fry (34.31%). The
number of fish examined were 180, 175, 160 and 160 for each age group, respectively.
The highest infestation with Trichodina sp. was found in spring (62.9%) followed by
winter (56.25%) then Autumn (41.37%) while the lowest infestation rate was in summer
(35%). A picture of heavily infected skin is shown in Fig. 2.
As indicated in the present study, the intensity of infection with Trichodina sp. was
generally low except few cases were fish mortalities have been observed in some ponds.
In light infestation, Trichodina sp. are usually present in small numbers of skin, fins and
160
Mohamed Abd El-Azez Hassan Hassan
FIG. 1. Trichodina sp. with peripheral adhesive disc followed by several circles of cilia and a central circle of
hooklets (Giemsa, × 400).
FIG. 2. Tilapia skin: showing a heavy infection by Trichodina sp. (Giemsa, × 100).
Trichodiniasis in Farmed Freshwater Tilapia...
161
gills of the infected fish and no clinical signs can be ascribed. On the other hand most
clinical signs and mortalities in some ponds were noticed in fish with heavy infections
and some cases of moderate ones due to exposure to one or several stress factors. These
factors may include, rough handling during transportation, overcrowdness, malnutrition,
high free ammonia (ranged between 0.8-2.16 mg/l) as well as low dissolved oxygen
(with average 1.4 mg/l in clinically infected ponds).
Clinical signs and postmortem examination revealed that diseased fish showed sluggish movement, loss of appetite, emaciation, loss of condition with larger head and
darker skin than normal. Some infected fish showed detached scales with pale skin
patches and more slimy skin. Erosion and ulcer on different parts of the body, blood
spots at the base of fins as well as laceration of membranous parts and sloughing of the
free portation of them were also observed in some infected fish. On the other hand excessive accumulation of mucous in the gill pouches and around gill filaments were frequently noticed in these infected fish. The gill filaments appeared hypraemic and swollen in some cases and pale in others. Signs of asphyxia including surfacing of fish,
gasping of air bubbles on the surface of water were also observed in heavily infested
fish. Finally they lost their normal swimming behaviour and died.
Histopathology
The skin of fish infected with Trichodina species showed, sloughing of the epidermal
layer and the remained dermis was oedematous and infiltrated with leucocytes and melanine-carrying cells which aggregate between the muscle layer (Fig. 3).
On the other hand, the epithelial lining cells of the gill filaments showed, proliferative changes, including, hyperplasia and hypertrophy. In most cases, the epithelial proliferation of the lamellae started from their apices and extended towards the basal portations. Sometimes, those proliferative changes extend to other lamellae where one, two,
or three lamellae appeared adherent to each other. In some fish, epithelial proliferation
was extensive leading to fusion of secondary lamellae (Fig. 4).
Therapeutic control
After 24 hour from the treatment with formalin (250 ppm), parasitological examination revealed that the Trichodina sp. were completely eradicated and most fish regained
their normal feeding response, mortality ceased, and fish began to appear healthy.
Discussion
Parasitic infestation of cultured fish in tropical and subtropical countries represents a
serious problem for aquaculture due to severe economical losses either as directly or indirectly (Roberts, 1978).
The data obtained from present study indicate that the Tilapia spp. have a variety of
clinical and post mortem changes from infection with Trichodina sp. Trichodina sp. are
commensals parasites on fish and have been reported in Tilapia (Abu El-Wafa, 1988;
El-Khatib, 1989; and Hassan, 1992).
162
Mohamed Abd El-Azez Hassan Hassan
FIG. 3. Skin section revealing loss of epidermis, disarrangement of collagen bundles and leucocytic infiltration of dermis with aggregation of melanine-carring cells (H & E, × 250).
Trichodiniasis in Farmed Freshwater Tilapia...
163
FIG. 4. Tilapia gills showing hyperplasia of interlamellar epithelium and hypertrophy of some secondary lamellae ( H & E, × 250).
164
Mohamed Abd El-Azez Hassan Hassan
It is also apparent in the present study that Trichodina spp. prevail on skin and gills
of Tilapia spp. Contrary to Lom (1962), who reported that the parasite occur only on the
skin in the fresh water, and on the gills in the marine forms. On the other hand, the
present findings confirm those of Snieszko and Axelrod (1971) in which they mentioned that Trichodina spp. occur on the gills and skin in numbers that could obscure the
normal structure of the epithelium.
The incidence of infestation with Trichodina sp. reached 49.6%. Such results are
nearly similar with that recorded by Okaeme et al., 1988 (50%) in cultured Tilapia spp.
in Nigeria. On the other hand higher rate of infestation of Tilapia spp. with the same
parasite was observed by Abu El-Wafa (1988) who recorded an infestation on 66% in
Egypt. However, lower results were obtained by El-Khatib, 1989 (30%) and Hassan,
1992 (12.5%). The present difference in the prevalence of infestation of the examined
fishes with Trichodina spp., from those previous workers, could be attributed to the different environmental conditions during the period of the present investigation and different locality.
Tilapia spp. of all ages and sizes, were susceptible to infestation with Trichodina sp.
The highest incidence of infestation was found in growing fish (66.6%) followed by fingerling (57.1%), then brood stock (50%) and lastly fry (34.3%). The present result conform to those of Paperna and Thurston (1968) who reported that Tilapia spp. of all ages
and sizes were susceptible to such parasitic infection throughout the year. It is worthy to
mention that infestation with Trichodina sp. varies between ages and sizes of Tilapia
spp. and can be attributed to overcrowdness and consequently changes to water quality
in ponds of growing and fingerling fish, compared with those of broods stock or fry.
Regarding the seasonal incidence, Trichodina infection was prevalent throughout the
year with maximum rate of infestation during spring season (62.9%), followed by winter (56.25%), then autumn (41.37%) and was lowest in summer (35%). The results
agree with the findings of McArdle (1984), Abu El-Wafa (1988) and El-Khatib (1989)
who reported that, Trichodiniasis was prevalent all over the year with maximum rate of
infestation during spring and winter.
It is also apparent in the present study that the intensity of infestation with Trichodina
sp. was generally low, excepting few cases where mass mortalities have been reported
in some ponds. In light infestation, fish do not show any signs of disease. However most
clinical signs and mortalities in some ponds were noticed in fish with heavy infection.
Mortalities in moderated infected fish is usually associated with one or more of stress
factors including, rough handling, overcrowdness, malnutrition, high free ammonia
(0.8-2.16 mg/l) as well as low oxygen (1.4 mg/l). Trichodina is an opportunistic parasite
which become pathogenic under stress full conditions (Ahmed, 1976; Paperna, 1980;
McArdle, 1984 and Eisa et al., 1985). On the other hand, Abdel Meguide (1989) reported that Trichodina sp. is a true and permanent parasite which was noticed to bring about
marked mortalities among newly-hatched grass carp.
The common lesions observed in the present study were in the form of black colouration, sloughing of scales with pale skin patches, laceration of membranous parts of fins
Trichodiniasis in Farmed Freshwater Tilapia...
165
and sometimes sloughing of free portion of them, erosion and ulcer in different parts of
the body were also observed and reported. These lesions are common in trichodina infection (Brown and Gratzek, 1980; McArdle, 1984; El-Khatib, 1989; and Hassan,
1992). In addition to these lesions, excessive accumulation of mucus on the skin and
gills was seen on infected fish. The massive production of mucus in case of Trichodiniasis infection is regarded as a defence mechanism to eliminate the parasite or dilute its irritating effects (Rogers and Gains, 1975).
The results of histopathological changes in skin and gills of Tilapia infested with Trichodina sp. revealed sloughing of the epidermal layer and the remaining dermis was
oedmatous and infiltrated with leucocytes and melanine carrying cells which aggregate
between the muscle layer and hypodermis forming a thick blackish band. This last finding may explain why the heavily infected fish had acquired characteristic dark colouration.
In addition to the above listed lesions, the epithelial lining cells of the secondary lamellae showed, proliferative changes, including, hyperplasia and hypertrophy resulting
in their fusion. This finding may interpret the behaviour of fish, heavily infested with
Trichodina sp., exhibiting distinct signs of respiratory disfunction.
With regard to parasitic control in the present study, inspection of the parasitised fish
(exposed to 250 ppm of commercial formalin, for one hour and examined after 24
hours) indicated that fish were free of the parasite. Mortality ceased and formalintreated fish appeared very healthy. Formalin is known to control ectoparasites in fish
(Ahmed, 1976).
In conclusion, present study emphasized the importance of keeping good husbandry
practice and favorable water quality parameters for successful rearing of fish. In case of
outbreak of trichodiniasis, formalin is an effective therapeutic agent.
Acknowledgements: First of all, prayerful thanks to our merciful God who gives me
every thing I have. My deepest gratitude and appreciation to Mr. F.A.A. Al-Jama, General manager of Fisheries Research Center, Al-Qateef, Eastern Province of Saudi Arabia
and and Mr. N.I. Fita, Technical Affairs Manager for their encouragement, and guidance which emboldened to carry out and fulfillment of this work. I wish to express my
sincere thanks to Prof. Dr. S.D. Deeb and all members of Pathology Department, Fac.
Vet. Med., Beni-Suef, Cairo Univ. for their supporting and facilities they offered while
conducting this work. Special thanks are to Mr. A.A. Al-Torif for his cooperation.
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