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Isolation and Identification of Aeromonas hydrophila and Saprolegnia sp. on Catfish (Clarias

gariepinus) in Floating cages in Bozem Moro Krembangan Surabaya

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2017 IOP Conf. Ser.: Earth Environ. Sci. 55 012038

(http://iopscience.iop.org/1755-1315/55/1/012038)

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Isolation and Identification of Aeromonas hydrophila and
Saprolegnia sp. on Catfish (Clarias gariepinus) in Floating cages
in Bozem Moro Krembangan Surabaya

Rahayu Kusdarwati1, Kismiyati2, Sudarno3, Hendi Kurniawan4 and Yudha Teguh Prayogi5
1
Department of Fish Health Management and Aquaculture Faculty of Fisheries and Marine,
Airlangga University, 031-5914151,
2,3
Department of Fish Health Management and Aquaculture, Faculty of Fisheries and Marine
4,5
Study Pograme Aquaculture Faculty of Fisheries and Marine, Airlangga University

[email protected]

Abstract. Catfish (Clarias gariepinus) is one of the familiar freshwater fish cultured in Indonesia
farmer. One of the reason is the high mortality of the catfish infected by Aeromonas hydrophila and
Saprolegnia sp. Motile Aeromonas septicemia (MAS) is a common bacterial disease, caused
by Aeromonas hydrophila, which affects freshwater fish. In Southeast Asia, the outbreak of this
disease was firstly reported from West Java in 1980, when a total of 82.5 tons a month of catfish
were lost, while in Central Java in 1984, the total loss was 1.6 tons. Saprolegniosis can cause
economic loss due to high mortality from its case reaching 10% to 50% . This research aimed to
identify and determine the percentage of A. hydrophila and Saprolegnia sp. the catfish farmed in
bozem Moro Krembangan, Surabaya, East Java. Meanwhile, a supporting parameter in this research
is the value of water quality parameter including pH, temperature, ammonia and dissolved oxygen
that were measured during sampling. The results showed that of the 20 samples taken from the two
cages, 19 fish were positively infected by A. drophila. percentage of infections of A. hydrophila that
infect umbo catfish in Moro Krembangan, was 95%, while the percentage Saprolegnia sp. was 90%.

1. Introduction

Surabaya has a system of outdoor water reservoirs that have the same functionality with reservoirs
namely Bozem. These Bozems include: Bozem Moro Krembangan, Kalidami, Bratang, Rungkut industrial,
Wonorejo, Kedurus and Broken Gorge. Ecosystem of Bozem is built as Surabaya is a city in flood-prone
with lack of water catchment areas. This is due to the development in the city of Surabaya that develops
rapidly. In General Bozem is built without cutting off rivers. In general Bozem serves as a controller and
water storage, and the society also utilizes it as a place to fish farming and irrigation of field (planning and
development agency of the city of Surabaya, 2010). One of the farmed fish in water of Bozem was catfish.
Catfish is one type of fish consumption that are quite popular in Indonesia. Some advantages of the fish are
to have meat that are thick and tasty as well as easy maintenance, its relatively quick growth, and high
selling prices, so that catfish arelargley farmed in to involves several factors: the surroundings
(environment), the farmed fish (host), and the disease-causing organisms (pathogens). A. hydrophila can
affect fish farming and often lead to outbreaks of disease with mortalities occasionally approaching 80-100
percent in a short period of 1-2 weeks [1]. While from the fungus genus, many Saprolegnia sp. infect eggs
and freshwater fish which can lead to massive death of fish in fish farms [2]. Saprolegnia sp. quickly
transmitted to other catfish that were in one pool. Thus, it spread more quickly and potentially inflicts
considerable losses for farmers [3].
Fish farming in Bozem Moro Krembangan having some problems such as the water environmental
problems that is less support, because the waters such a reservoir of water from various sources and disposal
of waste. Based on this research the reason they will do. The purpose of this research is to determine The
presence of infection Aeromonas hydrophila and Saprolegnia sp. on catfish (Clarias gariepinus) cultured
in Bozem Moro Krembangan Surabaya.

2. Research Methodology

2.1. Research Materials

Research materials that are used in this research are Clarias gariepinus obtained from Bozem Moro
Krembangan, Surabaya, aquades sterile, spirtus, alcohol 70%, selective medium Triptic Soy Agar (TSA),
Saboraud Dextrose Agar (SDA) and safranin. The instruments used are a petri dish, needle ose, mikropipet,
ring, light microscopy, object glass, hot plate stirer, thermometer, pH meters, DO the test kits and ammonia
test kit.

2.2. Research Methods

This study used a survey method. The determination of the sampling method used purpossive
sampling [4]. Sampling is to retrieve data of only a portion of the population who are expected to describe
the nature of the population from the object of research.

2.3. Work Procedures

2.3.1. Sampling
Catfish taken for the sample are consuming catfish with a size of 10-15 cm age 2-3 months with a
weight of 100-110 grams. The number of cages used for sampling amounted to two cages, the total
population is 400 catfish, in one population cage is 200 catfish, and taken a sample of 5% of the population
that is as many as 20 catfish out of a total population or 10 for each cage.

2.3.2. Sterilization of Tools

Equipment such as needles of ose before used first were sprayed alcohol 70% then conducted
burning directly. Petri dish was sterilized by autoclave at a temperature of 121 ° C with pressure 1 atm for
10-15 minutes.

2.3.3. Isolation of Aeromonas hydrophila and Saprolegnia sp.

The medium used for the isolation of A. hydrophila was Trypticase Soy Agar (TSA), while the
medium for isolation of Saprolegnia sp. was Saboraud Dextrose Agar (SDA). The isolation was carried out
using streak methods at medium of isolation. Furthermore, purification was done by moving the isolates
that grew into new media until pure isolate was obtained.

2.3.4. Identification of Aeromonas hydrophila and Saprolegnia sp.

Bacterial identification was conducted with the macroscopic test, Gram staining and biochemistry test.
Observations of macroscopic samples test of catfish included size, shape, and pigmentation seen from the
inside, beside and top. Microscopic tests were done by staining Gram by observing the color and shape of
bacteria as well as biochemical test was done. Biochemical test involved Motile test, indol test, O/F test,
and H2S test.
Identification of the fungus Saprolegnia sp. with macroscopic test and microscopic test.
Observations of macroscopic samples test catfish includde size, pigmentation (the colour of the colony) and
shape seen from the inside, beside and top. While the observations of the microscopic test were conducted
by observing the color, the shape of the hypha, forms spores, Spore and form layout cells of fungi by using
inverted microscope [5].

2.3.5. Calculating the percentage of Fish infected by Aeromonas hydrophila and Saprolegnia sp.
The percentage of infections was the number of fish infected by the bacteria/fungi that are divided
with many samples taken multiplied by one hundred. Resulting in the percentage of fish infected with the
disease.

Number of infected fish x 100

The percentage of catfish infected bacteria/fungi (%) = --------------------------------------
The number of fish samples

3. Results and Discussion

3.1 Identification and Isolation of Aeromonas hydrophila and Saprolegnia sp.
Isolation of A. hydrophila and Saprolegnia sp. was conducted on 20 samples of fish that were taken
that showed pathologic conditions including hemorrhagic septicemia and tail or fin rot. Isolation of a fungus
was conducted by taking body parts catfish that showed clinical symptoms of fungus infection. The fish
part isolated was part of the surface of the body, fins, gills, and tail, because the parts that would change as
result of infection of fungus Saprolegnia sp. On the skin of infected catfish there was smooth yarn
resembling cotton and spreading to the surface of the skin to another. This is in accordance with statement
[6], stating that fish infected with Saprolegnia sp. Have objects that resemble cotton on the surface of their
body, tail, fins or gills.
The results of the identification of the bacterial isolate showed negative staining Gram, motile,
form rod, while biochemical test results are fermentative, positive oxides, positive glucose and negative
indol.

a b

Figure 1. Bacterial isolates (a), (b) bacterial Morphology

Identification of fungus isolate in macroscopic colony characteristics can be seen with a yellowish
white with forms such as cotton. This is in accordance with [6], who states that the genus Saprolegnia have
characterized the colony resembling cotton, colored white and the colony can be found on the surface of
the skin, fins and gills.
Microscopically Saprolegnia sp. morphology appear to have long Hypha and aseptat (not
(not septad), tube-shaped, and has the zoospora on its corner. Sporangia widens and the short as
well as at the end of the sporangia looks a bit dark. The zoospora Saprolegnia sp was resulted from
the tip of the long Hypha . Zoosporangia Saprolegnia SP.that are long and cylindrical has a length
of 180-350 µm in width 20-24 µm [6]. Macroscopic and microscopically Observation can be seen
in Figure 2.

Figure 2. a colony of Saprolegnia sp.

One of the types of diseases that are often found in cultivated organisms is bacterial disease caused
by the bacterium Aeromonas hydrophilla, which is the pathogenic bacteria cause disease "Motile
Aeromonas Septicemia" (MAS), especially for freshwater fish species in tropical waters [7]. The bacterium
Aeromonas hydrophila is one of the dangerous disease-causing bacteria in freshwater fish farming. The
bacteria attack the Catfish that is one of the leading commodities in freshwater and can infect fish in all
sizes which can lead to death until it reaches 80%, resulting in huge losses in the freshwater fish farming
ventures [8].
Aeromonas hydrophila is a bacteria that can grow in the waters of the optimum operating
temperature of 20-30 ° C [9]. Aeromonas hydrophila is able to grow at temperatures up to 37 ° C [10].
Aeromonas hydrophila extracellular products of consist of the α and β hemolisin, protease, elastase, lipase,
cytotoksin, enterotoxin, gelatinase, caseinase, lecithinase, and leucocidin. The liver is the organ target of
Aeromonas hydrophila and disruption can have an effect on metabolic processes [11]. Aeromonas
hydrophila is pathogens because it only can cause disease in fish populations that are weak or as secondary
infections when fish are infected with other diseases [12].
The process of when Aeromonas hydrophila bacteria infects into the body of the fish is preceded
by that bacteria stick on the skin surface by utilizing the flagella to move towards the fish and then use
hooks to attach to strong on the outermost layer of a body of fish scales. Scales are covered by the substance
chitin. During the process of Aeromonas hydrophila bacteria takes plac, it produces enzymes that play a
role in kitinase layer of chitin degrades, so that bacteria can easily fit into the body of the fish. In addition
to utilizing the Aeromonas hydrophila kitinase, the bacteria also secrete enzymes such as lesitinase in an
attempt to get into the blood stream and go directly to the kidney to breed [13].
Saprolegnia sp. are fungi with yarn shape, resembling cotton, white to gray and Brown.
Saprolegnia sp. fungi belongs to the class of Oomycetes, also known as fungi with algae because its nature
is similar to that of algae but do not contain chlorophyll [14]. Compiled by the threads of a Hypha, it has
no bulkhead separators (aseptat), but the branched into misselium. The walls of Saprolegnia sp. Hypha are
composed of glycogen and cellulose [9]. Hypha branches Saprolegnia sp. form woven wool like yarn.
Saprolegnia sp. has a Hypha which is larger, tapered shape and able to spread faster than the other fungi.
3.2 The percentage of Clarias gariepinus Infected by Aeromonas hydrophila and Saprolegnia sp.
The data results of the counting of the Aeromonas hydrophila percentage of fungi farmed in floating
net cage in Bozem Moro Krembangan Surabaya can be seen in table 1.

Table 1. The results of the counting of the percentage of the Catfish infected Aeromonas hydrophila.
The number of fish infected by
The number of Fish Aeromonas hydrophila (fish) The percentage
Cages
Samples (fish) (%)
+ -
A 10 9 1 90
B 10 10 0 100
The total 19 1 95
20
number of

Results data of counting of percentage of fungi Saprolegnia sp. observed in cages floating in Bozem Moro
Krembangan Surabaya (seen in table 2)

Table 2. The results of the counting of the percentage of the Catfish infected with Saprolegnia sp.

The The number of fish

Cages of The number of fish infected by The percentage
floating sample (fish) Aeromonas hydrophila (fish) (%)
+ -
A 10 10 0 100
B 10 8 2 80
Total 20 18 2 90

On this study, the estimated value of water quality is obtained : temperature 30oC, DO 4 ppm, pH 6 - 7 and
ammonia 1.0 ppm.
Of the 20 fish samples taken, all were experiencing clinical symptoms such as wounds to the skin
also damaged fins and tail. Then at 19 fish’s kidney organ were suffered swelling and only 1 fish had not
experienced swelling in his kidney. The high percentage of fish infected by Aeromonas hydrophila and
Saprolegnia sp. was 95% and 90% indicating that the waters in Morokrembangan Surabaya Bozem were
less support for fish farming activities. Remember that waters had the function as shelter of water, such as
rain water and waste water, both household waste as well as industrial waste, with the aim to control the
occurrence of floods.
According to [15] Aeromonas hydrophila bacteria infection can happen in 4 levels, namely Acute
septicemia which is fatal, infection quickly with marked swelling of the internal organs. Sub acute, can be
seen with symptoms such as wounds and bleeding on the scales. Chronic can be seen with symptoms such
as ulcers and sores of the skin damage whose development lasts a long time. Latent, it can happen with not
showed symptoms of the disease, but on the internal organs contained disease-causing bacteria. While
infection of Saprolegnia sp. on fish generally are secondary infections that preced the onset of bacterial
infection, the aquatic environment are not heeded, the parasitic infestations, post-harvest handling, dense
stocking high causing easy fish infected with Saprolegnia sp. [16]. Bad water quality, low dissolved
oxygen, high organic matter content and the presence of bacteria will create an environment that is less
good for fish and cause stress. The low oxygen levels in the waters can cause fish become stressed out so
that the immune system can decline [17] . At that time, the pathogen will easily fit into the body of the fish,
either in the form of bacteria, fungi or parasites. While according to [18], pH changes can cause fish stressed
so that they are easily affected by desease, and automatically law pH can cause the damage on skin, so
pathogen can easily infect it. the value of optimal water quality for fish growth, namely: water temperature
28-32 oC. In addition to temperature, DO and pH, the content of ammonia is a factor to consider. The content
of DO is in the water so that fish growth is ideal not less than 2 ppm, pH 6.5 to 8.5 and an ammonia content
of less than 1 ppm.

4 Conclusion
Catfish (Clarias gariepinus) cultured in Moro Krembangan Surabaya, East Java, are mostly infected
with Aeromonas hydrophila and Saprolegnia sp., It is caused by the environmental conditions that are
unfavorable. The percentage of catfish (Clarias gariepinus) infected with Aeromonas hydrophila is 95%
and infected with Saprolegnia sp. is 90%.

References
[1] Lukistyowati, I. dan Kurniasih. 2012. Pelacakan Gen Aerolysin dari Aeromonas hydrophila pada Ikan Mas yang
diberi Pakan Ekstrak Bawang Putih. Jurnal Veteriner 13 (1) : 43-50.
[2] Akbar, J. 2008. Pemanfaatan Ekstrak Bunga Kecombrang (Nicolaia speciosa horan) Terhadap Penyembuhan
Infeksi Jamur Saprolegnia sp. Pada Ikan Nila Merah. Fakultas Perikanan Universitas Lambung Mangkurat.
[3] Khairyah U., Kismiyati., dan Kusdarwati R. 2013. Identifikasi dan Prevalensi Jamur pada Ikan Gurami
(Osphronemus gouramy) di Desa Ngrajek, Kecamatan Mungkid, Kabupaten Mungkid, Jawa Tengah.
[4] Singarimbun dan Effendi. 1995. Metode Penelitian Survei. Jakarta : PT Pustaka LP3ES Indonesia.
[5] Yuasa K., Novita P., Meliya P., dan Edy B.K, 2003. Teknik Diagnosis Penyakit Ikan Budidaya Air Tawar di
Indonesia. Japan International Cooperation Agency dan Panduan Diagnosis Penyakit Ikan. Balai Budidaya Air
Tawar Jambi. Jambi.Stoskopf, M. K. 1993. Fish Medicine. WB Saunders Company. Mexico.
[6] Rahmaningsih, S. 2012. Penagruh Ekstrak Sidawayah dengan Konsentrasi yang Berbeda untuk Mengatasi
Infeksi Bakteri Aeromonas hydrophyla pada Ikan Nila (Oreochromis niloticus). Jurnal Ilmu Perikanan dan
Sumberdaya Perairan.
[7] Sanoesi, E. 2008. Penggunaan Ekstrak Daun Pepaya (Carica papaya Linn) terhadap Jumlah Sel Makrofag pada
Ikan Mas (Cyprinus carpio L) yang Terinfeksi Bakteri Aeromonas hydrophila. Jurnal Penelitian Perikanan, Vol
11, No. 2.
[8] Indrawati, I. 2007. Potensi Ekstrak Rimpang Lengkuas (Alpinia galanga) terhadap jamur Saprolegnia sp. dari
telur ikan mas (Cyprinus carpio) Secara in Vitro. Skripsi. Fakultas Kedokteran Hewan. Universitas Airlangga.
Surabaya.
[9] Austin, B. and D. Austin. 1987. Bacterial Fish Pathogen. Ellis Hoorwood. London. pp. 250-262.
[10] Cipriano RC, GL Bullock and SW Pyle. 1984. Aeromonas hydrophila and Motile Aeromonad Septicemia of
Fish. Fish Diseases Leaflet 68, US. Fish and Wildlife Service. West Virginia.
[11] Kabata, Z. 1985. Parasites and Disease of Fish Cultureed in Tropics. Taylor and Francis, London.
[12] Mangunwardoyo, W., R. Ismayasari., E. Riani. 2010. Uji Patogenisitas dan Virulensi Aeromonas
hydrophila Stanier pada Ikan Nila (Oreochromis niloticus Lin.) melalui Postulat Koch. J. Ris. Akuakultur Vol.
5 Tahun 2010: 245-255.
[13] Nuryati, S., F.B.P. Sari., dan Taukhid. 2009. Identifikasi dan Uji Postulat Koch Cendawan Penyebab Penyakit
pada Ikan Gurami. Program Studi Budidaya Perairan. Fakultas Perikanan dan Ilmu Kelautan. Institute Pertanian
Bogor. Bogor.
[14] Sniezko SF and HR Axelrod. 1971. Diseases of Fishese. TFH Publication Ltd., Hongkong.
[15] Bruno D. W and Woo, P. T. K.. 1999, Fish Disease and Disorders vol. 3 (Viral, Bacterial and Fungal Infection).
CAB International Publishing. Wallingford, UK. Pp 366-432.
[16] Shafrudin, D., Yuniarti dan Setiawati, M., 2006. Pengaruh Kepadatan Benih Ikan Lele Dumbo (Clarias sp.)
Terhadap Produksi Pada Sistem Budidaya Dengan Pengendalian Nitrogen Melalui Penambahan Tepung Terigu.
Jurnal Akuakultur Indonesia 5(2) : 137-147.
[17] Taufik,P. 1984. Faktor Kualitas Air Dapat Mempengaruhi Timbulnya Suatu Penyakit Pada Ikan. Majalah
Pertanian No 3. Departemen Pertanian. Jakarta.