Journal of Fish Diseases 2011, 34, 801–804 doi:10.1111/j.1365-2761.2011.01282.

Short Communication

Arborescent organ necrosis syndrome in catfish, Clarias

gariepinus (Burchell): a case report

O O Oladele1, B E Olufemi2, G A Oladosu3, O L Ajayi4, A A Adediji1 and I O Arasi1

1 Poultry and Fish Diseases Diagnostic Laboratory, Animal Care Services Konsult (Nig.) Ltd Ogere Remo,
Ogun State, Nigeria
2 Department of Veterinary Medicine, Faculty of Veterinary Medicine, University of Ibadan, Oyo State, Nigeria
3 Nigerian Institute for Oceanography and Marine Research, Wilmot Point Road, Victoria Island, Lagos, Nigeria
4 Department of Veterinary Pathology, University of Agriculture, Abeokuta, Ogun State, Nigeria

Keywords: Arborescent organ, Aeromonas sobria,
catfish, Enterobacter cloacae, necrosis.
Previous reports have documented cracked head
disease (CHD) of catfish, which presents with
mortalities in fish with necrosis of the arborescent
organs and other pathologies, but the cause of the
disease was not completely understood. Affected
fish showed a reddish lateral line on the skull
between the two air chambers, possibly due to
haemorrhaging and necrosis of the osseous plate,
leading to cracks along the lateral line (Andras,
Shibabrata & Chowdhury 1992). In Nigeria, a case
study of CHD was carried out by Awa &
Alegbeleye (1991). Approximately 80% of the fish
were lost in that case including both sexes of African
catfish, Clarias gariepinus (Burchell), and both
potassium permanganate and formalinized alcohol
were used to treat/manage the disease.
The arborescent organ of Clarias allows it to
breathe air directly (Richard 2001). This implies
that necrosis of this organ will reduce the ability of
catfish to withstand low levels of dissolved oxygen
in water. Losses due to disease outbreaks come not
only from mortality, but also from cost of
treatment, loss of opportunity to sell and loss of
Correspondence O O Oladele, Poultry and Fish Diseases
Diagnostic Laboratory, Animal Care Services Konsult (Nig.) Ltd,
Ogere Remo, Ogun State, Nigeria.
(e-mail: vetmand07@yahoo.com)
growth during and after the outbreak (Omitoyin
2007).
In the case reported here, African catfish, weigh-
ing between 0.3 and 0.4 kg, were affected. They
were initially 10 000 in number and stocked in five
concrete tanks at 2000 fish tank at 125 fish per m2
before the outbreak.
Fresh carcasses and moribund samples of fish
were directly taken from the fish tanks. Clinical
signs and gross lesions were recorded. Some of the
harvested organs were preserved in 10% neutral
buffered formalin for histopathology.
The abdominal cavity of fish was opened and
the surface of the kidney was disinfected using
95% isopropyl alcohol. Swabs of the kidneys were
taken. An incision into the dorsal skeletal muscle
(caudal to the skull) was made, after disinfection of
the skin surface. Swabs of the exposed inner
portion of the skeletal muscle were taken. Liver,
spleen, milt sacs and arborescent organs were
seared before inocula from them were streaked on
sterile tryptone soy agar, MacConkey agar, TCBS
(thiosulphate–citrate–bile sucrose) agar and Sabou-
raud dextrose agar plates. Swabs of kidneys and
skeletal muscles were directly streaked on these
culture media, incubated aerobically at 37 °C for
24 h and later extended to 48 h for inocula on
Sabouraud dextrose agar.
Isolated bacteria were Gram stained. The Gram
reaction tests were confirmed using 3% KOH
solution (string test method) as described by
Gregersen (1978). Isolates were also subjected to

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 Journal of Fish Diseases 2011, 34, 801–804
oxidase and catalase tests. Other biochemical tests
were conducted (32 in total) on the bacterial
isolates using Gram-negative identification (GNID)
plates from Trek Diagnostic Systems as described
by Brimecombe, Grist, Butler, Chapin, Hall &
Knapp (2002). Each plate had its wells coated with
dry biochemical media, formulated for fluorometric
reading. Plates were incubated at 34 °C for 24 h
and then read using a SensititreÒ Autoreader.
Tissues from liver, skeletal muscle, kidney and
aborescent organs were fixed in 10% neutral
buffered formalin. These tissues were then dehy-
drated, embedded in paraffin wax, sectioned at
5 lm and stained with haematoxylin and eosin.
Isolates were subjected to antibiotic sensitivity
tests, using the disc diffusion method as described
by Baker, Breach & Leighton (1980). Antibiotic
discs (ofloxacin, augmentin (amoxycillin + clavula-
nate), tetracycline, nitrofurantoin, chlorampheni-
col, streptomycin, amoxycillin, gentamicin and
cotrimoxazole) were placed on the seeded surfaces
of nutrient agar plates. Streptomycin was adminis-
tered to fish with feed for 10 consecutive days at
20 mg kg)1 body weight.
The clinical signs observed were anorexia and
high morbidity (weak fish showed lateral recumb-
ence at the base of the tanks). Farm records showed
that mortality was very high over a period of
24 days before laboratory intervention. Percentage
mortality over the period was 72.8%.
Organisms isolated from this condition grew
only on tryptone soy agar and MacConkey agar. On
MacConkey agar, one of the isolates had colonies
that were pink, discrete, low convex and entire,
measuring about 1–2 mm in diameter. It produced
Table 1 Results of biochemical tests on isolated bacteria using SensititreÒ Gram-negative identification plates
Isolate 1
Enterobacter cloacae
96.74% probability
FR1 –ve; Urea –ve; Ornithine +ve; Sorbitol +ve
Xylose +ve; FR12 +ve; Sucrose +ve; FR9 +ve
FR3 +ve; Trehalose +ve; FR8 –ve; Mannitol +ve
Maltose +ve; FR4 –ve; Inositol –ve; FR10 +ve
FR5 –ve; Fructose +ve; Aesculin +ve
Arabitol )ve
Arabinose +ve; Lysine +ve; TDA )ve
Raffinose +ve
FR7 +ve; Arginine +ve; FR6 +ve; Cellobiose +ve
Malonate +ve; Pyruvate +ve; Citrate +ve; Agmatine +ve
Biocode 3FD96DFF
FR1, Lysine AMC (0.12 g L ); FR3, Phosphate MU(0.4 g L ); FR4, a-D-Glucoside MU(0.2 g L)1); FR5, Proline AMC (0.06 g L)1); FR6,
)1
a-d-Galactoside MU(0.5 g L)1); FR7, c-Glutamine AMC (0.03 g L)1); FR8, Bisphosphate MU(0.19 mg L)1); FR9, b-d-glucuronide MU(0.19 g L)1);
FR10, b-d-galactoside MU (0.19 mg L)1); FR12, 2-acetamido-2-deoxyglucoside + a-l-arabinoside MU(0.5 g L)1).
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O O Oladele et al. Arborescent organ necrosis in catfish
a slight greenish metallic sheen on eosin methylene
blue (EMB) agar, but was citrate positive on
SimmonÕs citrate agar and was oxidase negative.
The GNID test identified it as Enterobacter cloacae.
The second isolate, which was oxidase positive,
grew as a weak lactose-fermenting organism (very
light pink-coloured colonies) on MacConkey agar
with the colonies measuring between 0.5 and 1 mm
in diameter. It was identified as Aeromonas sobria by
the GNID test (Table 1). Both organisms were
Gram-negative short rods and were catalase
positive. Both isolates were sensitive only to
streptomycin.
During post-mortem examination, depigmenta-
tion of the skin, inflammation of the dorsal muscles
just caudal to the skull, opacity of the lens, pale gills
with some being ash-coloured and necrotic arbo-
rescent organs covered with mucoid exudates were
observed. A grey and putrefying fascia-like mass was
Figure 1 Necrosis of the arborescent organ of Clarias gariepinus
with the presence of mucoid exudates.
Isolate 2
Aeromonas sobria
100% probability
FR1 –ve; Urea –ve; Ornithine +ve; Sorbitol )ve
Xylose –ve; FR12 +ve; Sucrose +ve; FR9 )ve
FR3 +ve; Trehalose +ve; FR8 +ve; Mannitol +ve
Maltose +ve); FR4 –ve; Inositol –ve; FR10 +ve
FR5 +ve; Fructose +ve; Aesculin –ve; Arabitol )ve
Arabinose –ve; Lysine +ve; TDA –ve; Raffinose +ve
FR7 –ve; Arginine +ve; FR6 +ve; Cellobiose )ve
Malonate –ve; Pyruvate +ve; Citrate +ve; Agmatine +ve
Biocode 26F9C567
)1
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Figure 2 Presence of a grey fascia-like mass on the arborescent
organ of Clarias gariepinus.
Figure 3 Goblet cell hyperplasia and submucosal haemorrhage
with haemosiderosis in the arborescent organ of Clarias gariepi-
nus.
also observed on the necrotic arborescent organs
(Figs 1 & 2). All kidneys and spleens were
congested, and the liver samples had a light to
dark brown colouration, some with a slight greenish
taint.
Histological sections of the arborescent organ
revealed goblet cell hyperplasia and submucosal
haemorrhage with haemosiderosis (Fig. 3).
Aeromonads are very commonly reported as
pathogens of fish. However, Sekar, Santiago, Vija-
yan, Alavandi, Raj, Rajan, Sanjuktha & Kalaimani
(2008) demonstrated the involvement of E. cloacae
in the mortality of mullet. Species of Enterobacter
were also found to be associated with mortality in
reared channel catfish (Sherman, Peter, Crosby,
James, MacMillan & Robert 1992).
In the case reported here, treatment was carried
out orally for 10 consecutive days with streptomy-
cin administered at 20 mg kg)1 body weight.
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O O Oladele et al. Arborescent organ necrosis in catfish
Cumulative mortality over 24 days was 6508 fish
with an average daily mortality of 271 fish, while
during the treatment period, total mortality was
774, with an average daily mortality of 77 fish. A
day after the end of treatment, only two fish were
found dead. Percentage mortality over the study
period was 72.8%.
Streptomycin is an aminoglycoside and said to be
poorly absorbed from the gut (Brander, Pugh,
Bywater & Jenkins 1991), but it showed good
efficacy in this case.
Control of this outbreak also involved decon-
tamination of the environment as it was noticed
that mortality did not abate in tanks where fish were
infected. Fish from affected tanks were transferred
to thoroughly washed, disinfected and rinsed tanks.
Formalin (38%) was used at 50 mL L)1 of water to
disinfect empty tanks, which were later rinsed and
used for treatment.
Apart from the case reported here, consultation
with other affected fish farmers indicated a strong
link between these high and unusual mortalities
and the quality of the fish meal used in formu-
lating fish feed which always had very high level of
coliforms and non-coliforms as shown by micro-
bial analysis. This suggests an urgent need for
quality assurance of inputs in fish farming, which
should be the priority of regulatory bodies and fish
farmers and will help prevent a collapse of the
aquaculture industry.
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Received: 27 December 2010
Revision received: 25 March 2011
Accepted: 25 March 2011