Egy. J. aquac., Vol. 4, No.

(2):1-12 (2014) ISSN: 2090-7877

Causes of mortality in Discus fish (Symphysodon) and trials for

treatment
Nadia A. Abd El-Ghany, Nahla R. El-khatib, and Soad S. A. Salama
Fish Diseases Department- Animal Health Research Institute, Dokki, Giza.

Abstract
Discus (Symphysodon) from the family cihildae is one of the most
popular and expensive aquarium fish. In the past few years, farming of
this fish has been well developed in Egypt. In this study, a total number
of 30 Symphysodon spp collected randomly from private freshwater
ornamental fish farm in Kalubia Governorate at December 2013. Discus
fish suffered from mortality after the onset of anorexia, eye cloudiness,
ascites, excessive body mucus, frayed dorsal fin and tail rot. They were
subjected to clinical, postmortem, parasitic, bacterial and mycotic
examinations to clarify the causative agents of mortality. The recovered
fungi were Fusarium solani, F. oxysporum and F. moniliform with the
prevalence of 50, 33.34 and 16.66% respectively. Bacteriological
examination revealed that isolation of Aeromonas hydrophila with
prevalence 60% and it was sensitive to Ciprofloxacin. The parasitic
examination revealed that fish were infested with Spironucleus spp with
prevalence of 80%. The Symphysodon spp which were concomitantly
mixed infected by bacterial (A. hydrophila), fungal (Fusarium) and
parasite (Spironucleus) were subjected to trials for treatment either by
chemical or physical.
Key words: Discus fish, Fusarium spp, Aeromonas hydrophila, Symphysodon
spp, Spironucleus spp, treatment.

Introduction and poor management. At present, various

Ornamental fish keeping has become
an ever more common hobby global.
Several species of ornamental fishes are
imported from Southeast Asian countries
into other countries (Iqbal et al., 2012).
Discus is one of the most popular and
expensive aquarium fish and considered
as a difficult-to-culture species due to its
complex reproductive behavior, involving
parental care of eggs and newly hatched
larvae (Chelappa et al., 2005 and
Manochehr et al., 2014)
Infectious diseases pose a constant and
serious threat to ornamental fish that are
farmed intensively under conditions of
high stocking density, poor water quality
diseases have been reported in ornamental
fish including bacterial diseases (Pate et
al., 2005), viral diseases (Hedrick and
McDowell, 1995) and parasitic diseases
(Leibowitz and Zilberg, 2009 and Smith
and Roberts, 2010).
Discus fish require higher water
temperatures than most other tropical
aquarium fish, so they should be
maintained between 28ºC – 31ºC
Giovanetti and Lucanus (2001).
Temperature lower than 27ºC caused
Discus became significantly more
susceptible to diseases.
In Egypt, There are few studies on
diseases of discus and causes mortality.
1
 Causes of mortality in Discus fish (Symphysodon) and trials for treatment
The objective of this study was to survey
the causes of loss of such fish in
propagation farm, trails for treatment and
management control.
Materials and Methods
Naturally infected Fish
A total number of 30 Symphysodon
spp of range body weight (0.5 - 3 g)
suffered from skin lesions and anorexia.
They were randomly collected from
private freshwater ornamental fish farm in
Kalubia Governorate (complained from
Discus mortality) at December 2013 and
transported alive to Fish Diseases
research department Lab Animal Health
Research Institute. Fish hold in fully
prepared aquaria for observation and
examination as soon as possible in same
farm water.
Clinical and postmortem examinations
All of the collected fish were
subjected to clinical signs and
postmortem findings according to (Plumb
and Bowser, 1983).
Mycological examination
The mycological methods used in this
investigation were described by Chinain
and Vey (1988). Tissue specimens
measuring approximately 5-10 mm were
taken under aseptic precautions from
liver, spleen, kidney, gills and skin and
inoculated into plates containing
Sabouraud's dextrose agar medium with
500 units' penicillin and 2 mg
streptomycin per ml-1 to prevent bacterial
growth according to (Benk and Rogers,
1970).
Cultures were kept at 25oC for 1-4
days. Pure cultures were established using
single spore isolation method a small part
of fungal mycelium from the 1-4 days
culture was transferred using sterile
medical needle and inoculated on PDA
2
(Potato Dextrose Agar, (Booth, 1971)).
Cultures were incubated at 25oC for 3-7
days. Post inoculation, the spores were
stained with lactophenol cotton blue and
examined microscopically.
A morphometric identification of the
fungal isolates was carried out based on
culture characteristics, namely colony
colour, type of mycelium, shape and
septation of micro and macroconidia
(Burgess, 1981).
Bacteriological examination
Samples from liver, spleen, kidney,
gills, ascetic fluid and gas bladder of
examined Symphysodon spp were
streaked onto nutrient agar, trypticase soy
agar, Rimler- Shotts medium (RS) and
thiosulphate citrate bile salt agar (TCBS)
plates then incubated at 25ºC for 24-48
hrs. The growing colonies were picked up
in pure form and reinoculated into
trypticase soy agar for further
identification.
Isolates were identified by cultural,
morphological and biochemical
characterization according to Austin and
Austin (2007) and through using
commercial miniaturized API®20E
system (Biomerieux, France) according to
manufacturer’s instructions.
Antibiogram
It was carried out using various
chemotherapeutic agents against the
isolated A. hydrophila and judgment of
the obtained results in comparison to
interpretive standards was applied as
described by (Quinn et al., 2002).
Parasitological examination
Fish were examined externally and
internally for gross signs of parasitism
and any abnormalities. Smears made
from skin lesions and from various sites
along the gastrointestinal tract and of the
blood, and imprints made of internal
 Nadia et al.,
organs (liver, kidney, spleen, and gall
bladder). Microscopic examination of wet
mounts under bright field and phase
contrast X40. Stained smears with Giemsa
stain were used in detailed examination
and identification according to (Kent et
al., 1992 and Buchmann and Uldal,
1996).
Treatment experiment
A total number of 60 natural infected
Symphysodon spp. with mixed bacterial,
mycotic and parasitic infection were
divided into 6 groups, ten fish each. The
fish were holding in six full glass aquaria
40×40×50 cm with Chlorine free tap
water. Fish starved two days before
treatment according to (Innes, 1966). All
fish were observed over two weeks period
for morbidity and mortalities and
examined 24 hours after the end of the
treatment. A treatment was considered
effective when it caused a complete
removal of infection of all fish used in the
assay (Tojo and Santamarina, 1998).
Chemical treatment
The natural mixed infected
Symphysodon spp. in group one were
treated by exposed long bath treatment of
Metronidazole 5ppm for 12 hour for
three times day after day and 50% water
changes before drug administration. The
fish in group two were exposed to
commercial antibiotics ciprofloxacin
(5ppm for 5 days and 50% water changes
before drug administration). The choice of
these drugs was according to sensitivity
test result and the doses according to
(Noga, 2010).
All chemical treatments were applied
at water temperature of 20°C and
dissolved oxygen was censured by regular
aeration.
Physical treatment
In this type of treatment, there were
two aquaria ( group three and group four)
contain natural mixed infected Discus fish
held at 25°C and 30°C using
thermostatically control water heaters.
Control infected Discus fish groups were
subjected to similar circumstances, but
without use of any chemicals.
Results
Clinical signs
Naturally infected Symphysodon spp
showed ulceration on the skin especially
on the head and dorsal fins. Fish suffered
from exophthalmia, eye cloudiness,
congested gills abdominal distention,
excessive body mucus, frayed dorsal fin
and tail rot Fig .1 (A, B & C).
Post- mortem findings
Postmortem findings of naturally
infected fish revealed congestion of
visceral organs, in some cases paleness
and focal liver with yellowish nodules,
enlargement of spleen, yellowish fluid in
abdominal cavity. The intestine showed
yellowish mucus and haemorrhages of gas
bladder. In some cases gas bladder filled
with yellowish fluid Fig .1 (D).
Fungal study
The recovered fungi isolated on
Sabouraud's dextrose agar medium and
preliminarily identified on PDA were
identified morphometrically as members
of the genus Fusarium, namely Fusarium
solani (50%), F. oxysporum (33.34%) and
F. moniliform (16.66%). The prevalence
of recovery of such Fusarium species in
infected ornamental fish Symphysodon
spp reached (100%) (Table, 1).
Morphological criteria of Fusarium
species were summarized
Fusarium solani: colony grow
rapidly on PDA with dense aerial white to
3
 Causes of mortality in Discus fish (Symphysodon) and trials for treatment

cream mycelium in concentric rings, characteristic of bacterial isolates are

abundant, floccose, somewhat powdery in summarized in (Table 2) and showed that
aged cultures which reverse to brownish the isolates were identified as A.
colour, showing average growth rate per hydrophila.
day at 25oC Fig. 2 (B). Microconidia
In-vitro sensitivity tests of isolated
usually abundant, ellipsoidal, fusiform or
bacterial strain A. hydrophila to different
kidney shaped after 2-3 days in fresh
chemotherapeutic agents revealed that the
isolates. Macroconidia were abundantly
isolates were sensitive to Ciprofloxacin.
developed after 4-7 days with thick
walled having 3 to 4 septa, straight, Parasitological study
parallel sided for most of length. The
There were no ectoparasites detected
apical cell blunt had rounded end Fig. 2
and only diplomonad flagellates
(C & D).
Spironucleus spp in intestine, liver, gall
Fusarium oxysporum: colonies are
bladder, spleen, heart, skin and fins was
dense and floccose, white mauve
mycelium, which reverse dark brown. observed under a light microscope and
Microconidia were long produced in false phase contrast X40.
The intensity revealed that higher
heads, carried by short conidiophores and
occurrence in the hind part of the intestine
consisted of one or two cells,
than internal organs. The prevalence of
chlamydospores with a thick wall,
parasitic infestation in Symphysodon spp
produced singly or in pairs. Macroconidia
only slightly curved, usually with 3 septa, was 80% (Table, 1). The morphological
occasionally more, thin walled with a characteristics of diplomonad flagellates
under light microscopes indicated that the
notched or foot-shaped basal cell and
parasite was Spironucleus Spp.
short.
The morphology of the (n= 20 )
Fusarium moniliform: colonies are
dark violet with dense aerial mycelium trophozoite stages have transparent
which is delicately floccose to felt. elongated pyriform body and consider
smallest diplomonad flagellates, with a
Microconidia were formed in chain,
body range of 13- 20 mm long and 5.5-
fusiform to clavate with a slightly
10.4 mm wide. The parasite is anterior
flattened base, they occasionally become
tapering and intertwined elongate
one septate. Macroconidia are
inequilaterally fusoid, delicate, thin compact nuclei S-shaped and wrap around
walled, often sharply curved apical cell each other at their narrow anterior ends.
The body is emerging three anterior
and pedicel at basal cell, they are 3-7
flagella, and one posterior flagellum (Fig.
septate.
2 (A)).
Bacteriological study
Treatment trials
Bacteriological examination of liver,
spleen, kidney, gills, ascetic fluid and gas In the present investigation, the trials
bladder of examined fish revealed that 72 to treat natural mixed infected
Symphysodon spp. either chemical or
bacterial isolates were recovered and
physical. Table (3) shows that all mixed
characterized from 18 infected fish and
infected fish in group one which were
the total prevalence of bacterial Infection
was 60% (Table, 1). Phenotypic treated with chemical and physical
management (Metronidazole 5ppm for 12
4
 Nadia et al.,

hour for three times, day after day and
50% water changes before drug
administration) have recovered from
infection within 2 weeks post treatment
with complete disappearance of clinical
signs meanwhile all mixed infected fish in
group two which were treated with
ciprofloxacin (5ppm for 5 days and 50%
water changes before drug administration)
have not completely recovered from
infection, and two weeks post treatment
morbidity rate was 50% of fish .
Table (4) shows that all mixed
infected fish that exposed to physical
management in the form of increase of
water temperature to 30°C were recovered
from infection within two week post
exposure but 25°C not completely
eradicated infection, however there was
some fish loss in compared with chemical
treatment by Metronidazole include
physical management in the form of
partial (50%) water changes.
On the other hand, all of control
mixed infected fish in group three and six
were dead.

Fig. (1): show naturally infected Symphysodon spp suffering from ulceration on the skin especially on the
head, dorsal fins, tail rot and ascites. Arrow (A, B & C) show severe congestion of internal organs. Arrow (D).

5
 Causes of mortality in Discus fish (Symphysodon) and trials for treatment

Fig.2. show Giemsa stained Spironucleus isolated from Symphysodon spp. X100, arrow(A).
Colonies of Fusarium solani on PDA showing aerial, white to cream mycelium in concentric rings (B).
Microconidia of Fusarium solani after 2-3 days stained with lactophenol cotton blue showing fusiform shape.
(X40) arrow(C). Macroconidia of Fusarium solani showing slightly curved, more and thin walled (X 40)
arrow (D).

Table (1): The Prevalence of natural mixed infection of examined Symhysodon spp.

No. of examined No. of naturally

Types of infections %
Fish infected Symhysodon spp
Fungal infection
30 100
Fusarium sp.
Parasitic infection 30
24 80
Spironucleus
Bacterial infection
18 60
A. hydrophila

6
 Nadia et al.,

Table (2): Morphochemical characteristics of bacterial isolates from Symhysodon

spp.

Morpho-chemical Morpho-chemical
A. hydrophila A. hydrophila.
tests tests
Motility + Indole +
Growth on R.S + Gelatin liquefaction +
Growth on
- Gas from glucose -
MacConkey`s
Ferementation of
Cytochrome
+ glucose – +
Oxidase
Citrate - Esculin hydrolyze -
Growth at: 4°C - Gram stain -
Growth at: 41°C - Swarming -
Fluorescent
- H2S -
Pigments
Arginine
- Nitrate reduction -
Dihydrolase
Catalase + Urease -
Assimilation of :
Assimilation of :
Sucrose -
Mannitol -
Sorbitol -
Maltose -
Glucose +
Rhmnose -
Arabinose -

-: mean negative results, +: mean positive results

Table (3). The cumulative mortality rate and the morbidity rate of treated
Symhysodon spp by chemicals treatment.

Time in Control Metronidazole Ciprofloxacin

weeks
Cumulative Cumulative Cumulative
Morbidity Morbidity Morbidity
Mortality Mortality Mortality
% % %
% % %
0 0 100 0 100 0 100
1 50 50 10 20 20 60
2 100 0 10 0 30 50

7
 Causes of mortality in Discus fish (Symphysodon) and trials for treatment

Table (4). Cumulative mortality and morbidity rates of treated Symhysodon spp by
physical treatment (heat)
Control Water temperature Water temperature
Time 20ºC 25ºC 30ºC
in Cumulative Cumulative Cumulative
Morbidity Morbidity Morbidity
weeks Mortality Mortality Mortality
% % %
% % %
0 0 100 0 100 0 100
1 50 50 10 80 0 20
2 100 0 30 50 0 0
N.B.: The number of fish in each group is 10 fish

Discussion affect fish are ulcerative skin lesion,

Ornamental fish keeping has become lethargy, abdominal distension, ascites
an ever more common hobby global. and anorexia.
Discus is one of the most popular and
Regarding the causative agents of
expensive aquarium fish and considered
mortalities among Symphysodon was the
as a difficult-to-culture species. Studies
mixed infection with A. hydrophila in
on diseases of ornamental fishes
prevailing of 60%, Spironucleus spp 80%
especially Discus in Egypt is few. Studies
and Fusarium spp 100%. These results
on external parasites of ornamental fishes
are consistent with those obtained by
were done more frequently than the ones
(Pate et al., 2005 and Smith and
on internal parasites.
Roberts, 2010).
In the present study, causes of
Several cases of infection related to
mortality among Discus (Symphysodon
Fusarium spp. have been reported in
spp) fish were investigated at late winter
ornamental fishes farm (Manochehr et
of 2013. Naturally infected Symphysodon
al., 2014). Species of Fusarium
spp showed ulceration on the skin
responsible for the disease in ornamental
especially on the head and dorsal fins.
fishes have been identified F. solani and
Fish suffered from exophthalmia, eye
F. moniliform (Iqbal and Ramsha,
cloudiness, ascites, excessive body
2013). F. oxysporum causes several plant
mucus, frayed dorsal fin and tail rot.
diseases Burgess (1981) and a gill disease
These results were in agreement with
in the crayfish (Maestracci, 1987). The
Paull and Matthews (2001) who showed
fungus species is thus found in plants and
that the infected cichlids with
in different species of freshwater and
Spironucleus vortens, showed varied
marine crustaceans.
clinical signs from external to severe
Fungal identification was based on the
systemic symptoms and disagreed with
presence of fungal hyphae and conidia by
Kulda and Lom, (1964) showed that
using light microscopy. Hyphae of F.
infected angelfish had no signs of the
solani appear septated and the
disease although they were infected by
characteristic canoe-shaped macroconidia.
Spironucleus elegans. Also, Roberts et
These results agreed with previously
al. (2009) found that the most common
described (Lightner, 1996).
clinical signs of motile aeromonads that
8
 Nadia et al.,
Also, the morphological culture and
biochemical character of isolated A.
hydrophila was similar to recorded by
(Awad, 2011).
Control of fish disease is currently
based almost entirely on chemotherapy
and it will entirely retain a role in the
management of fish culture systems
(Roberts, 1995).
Regarding to the sensitivity of A.
hydrophila to different antimicrobials, the
present study showed that Ciprofloxacin
was the drug of choice. Similar results
were recorded by Musa et al. (2008) and
Awad (2011) who mentioned that
Ciprofloxacin has proved a world-wide
efficacy to control or treat bacterial
infections in fish. Successful control of
bacterial diseases of cultured fish is a
major management and economic
problems for the aquaculture industry
(Plumb, 1999).
The results of the treatment indicated
that chemical treatment with
Metronidazole included physical
management of partial water changes was
effective to overcome parasitic and
bacterial infection and absence of
morbidity and mortalities after two weeks.
These results agreed with Gratzek,
(1983) and Woo (2006) who mentioned
that Metronidazole is an antiprotozoal
medication used in fish, although
Sangmaneedet and Smith (1999) found
that combination of metronidazole and
Magnesium sulfate have been the most
effective chemotherapeutic agents in
inhibiting the growth of S. vortens in
angelfish but the physical management of
change water may help for removing
such parasite before infecting another
fish.
In the present study, physical
management in the form of increase of
water temperature to 30°C showed no
mortality and overcome all infections but
at 25ºC not completely disappeared of
morbidity and mortality rate. These
results are consistent with those obtained
by Sung and Cook (1981) for increasing
temperatures in the range of 15-35°C
decrease the potential for growth
Fusarium spp. and Kersters et al. (1995)
who investigated the incidence of
Aeromonas hydrophila is found in high
numbers when the temperature was
around 20–25 ºC and were rarely detected
during cold seasons. Also obtained results
supported those reported by Bassleer
(1983) and Buchmann and Uldal (1996)
who found the completion of Hexamitid
growth was at temperature15ºC-20ºC and
the optimal performance at 10ºC .
Gratzek (1983) and Sangrnaneedet
and Smith (1999) who reported that the
rise in water temperature up to 28ºC-29ºC
enhance the fish’s immune response to
overcome diseases occurrence.
In conclusion, the causative agents of
Discus mortalities were concurrent
parasitic / bacterial and fungal infections.
This circumstance was the real scenarios
for infections in the fish environments
where water is naturally inhabited by the
ubiquitous bacteria, fungal and parasites
and any stress factors causing fish
diseases. In case of rearing tropical fish
(Discus) we need to take into our
consideration the special nature of such
fish. So physical management and
chemical treatment should be applied if
disease establish.
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parasitic diseases of rainbow trout
Oncorhynchus mykiss. I: Hexamita
salmonis. Dis Aquat Org., 33:51-56.
11
 ‫‪Causes of mortality in Discus fish (Symphysodon) and trials for treatment‬‬

‫‪Woo, P.T.K. (2006). Fish Diseases and‬‬ ‫‪Metazoan‬‬ ‫‪Infections,‬‬

‫‪Disorders, Volume 1: Protozoan and‬‬ ‫‪2nd edition, CABI, U.K.‬‬

‫الملخص العربى‬

‫أسباب النفوق فى األسماك القرصية ومحاوالت للعالج‬

‫نادية احمد عبد الغنى ‪ -‬نهلة رمزى الخطيب و سعاد صبرى عبد الحليم‬
‫قو با أمرا األسماك‪ -‬مع د با صاة الاي ا – الدق ‪ -‬الجيزم‬

‫(سنيمى سن ن ) هن ادندم من أسنماك الزيننة‬ ‫تعتبر أسماك الدسكس (القرصية) من ون‬

‫ا ت ننا‪ ,‬ع ن الوننن اق القليلننة الما ننية ت ن ت ن ير‬ ‫الجميلننة األكث نر شننعبية لكن ننا مكلىننة ع ن‬
‫استز ا هذه االسماك بشكل جيد ع مصر‪.‬‬
‫ع هذه الد اسة ت تجمين ندن‪ 03‬سنمكة شن اييا من أسنماك الدسنكس (سنيمى سن ن )‬
‫م مز ة أسماك ينة خاصة عن مااعةنة القلي بينة عن نيونمبر ‪ 3300‬كاون األسنماك تعناو‬
‫أمننرا األسننماك ‪ -‬مع نند بان صنناة‬ ‫من ووننبة وىن ة اليننة تن ا سننال ا الن معمننل بان‬
‫مل عا صاق ميكر بيل جية طىيلية‪.‬‬ ‫الاي ا ت عاص ا عاص ظاهرى عاص تشريا‬
‫كاو العالماق المر ية ل األسماك قبل النىن ة تبندأ بىقندا عن الشن ية‪ ,‬تامنة عن العني ‪,‬‬
‫ي نانم ع ن كميننة المانناط المج ن للجو ن ‪ ,‬تعى ن ع ن الز نىننة الة ريننة الذيليننة ج ن ن س ن ايل‬
‫ا تشادية نم ية ع التج يف الب ن ث النى ة‪.‬‬
‫أ ننا الىا ن ط ال ىيليننة البكتيريننة الى ريننة أ األسننماك المصننابة قنند ت ن ننز من ننا‬
‫موننبباق مر ننية ميكر بيل جيننة طىيليننة كاونن كنناألت ‪ :‬الى رينناق المعز لننة تنتمنن لى ننر‬
‫عي ا يننن و سننن الو (‪ , )٪03‬ع نننر الىي ا يننن و أ كويونننب ي (‪ )٪00.03‬الىي ا يننن و‬
‫م ويلى ن و ‪ .٪01.11‬كشننف الىاننص البكتري ل ن ج ن ننز تننرم اير م ونناو هينند عيال‬
‫(‪ )٪13‬جد او ا دواسة للويبر عل كواسي ‪ .‬كاو االصابة ال ىيلية ب ىينل سنبير ويكلس عن‬
‫األمعاء األ ضاء الداخلية بنوبة (‪.)٪03‬‬
‫قننند أ نننا الد اسنننة أ االصنننابة الميكر بينننة كاوننن ماتل نننة ببكتيرينننا (اير م وننناو‬
‫هينند عيال)‪ ,‬الى رينناق (الىي ا ي ن و) ال ىيلينناق (سننبير ويكلس) قنند ت ن الننتاك ع ن األصننابة‬
‫المر ية ب رة العالج الماتلىة إما الكيميايية أ استاداو الارا م ‪.‬‬

‫‪12‬‬

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