APLICATION OF PROBIOTIC AND MOLASES FOR IMPROVING OF REARING
MEDIA ON CRABLET MASS PRODUCTION OF SWIMMING CRAB (Portunus
Sugeng Raharjo2, Nana S.S Udi Putra2, Eddy Nurcahyono2, Makulau Sultan2
E-mail : firstname.lastname@example.org
Center for Brackhiswater Aquaculture Development Takalar
Bontoloe - Galesong, Distric of Takalar South Sulawesi 92245 Indonesia
An environmental friendly solution for improving water quality of rearing media of swimming crab
hatchery (Portunus pelagicus) used probiotic bacteria. The bacteria needs the spesific substrate for
growth well. Molases is a good substrate and carbon source for bacteria growth and develop, so should
be conducted the study to know the effectiveness applied method. Results showed that application of
the probiotic bacteria within molases in water media of swimming crab larvae (Portunus pelagicus,linn)
tend to increase the survival rate, body weight, lenght of carapace, and able to accelerate the growth of
zoeae stage, in addition also reduce the population growth of Vibrio sp (102 cfu/ml). Application of
probiotic bacteria and molases could accelerated the growth stage from larvae to crablet (megalop at 9 th
day and crablet at 14th day) were more faster than only using the probiotic bacteria (megalop at 10th day
and crablet at 15th day) and also for weight and lenght of carapace in the end of stage, but that not
significantly consistant different between treatments. Total Bacteria population in treatment of probiotic
bacteria and molases is about 4,2 X 106 CFU/ml and Vibrio sp is about 2 X 102 CFU/ml, contrary for
treatment bacteria only is about 6 X 104 CFU/ml (Total bacteria) and Vibrio is about 4,2 X 103.CFU/ml.
Furthermore molases was effective in improving of rearing water media on mass producing crablet of
Key words: Probiotic bacteria, molases, swimming crab
Hatchery of swimming crab (Portunus pelagicus,linn) plays an important role in swimming crab
seed supply to aquaculture activities in the farmers pond. The constraints often found in hatchery which
survival rate was unstable, despite of the achieving of the survival rate until crablet-10 was about
45,68% (Nurcahyono, et al, 2008). The main problem of lower survival rate is due to presence of
pathogen bacteria in water media while water quality was suddenly dropped. In several seasons water
source were bloomed by pathogen bacteria mainly Vibrio sp. Several treatments were conducted for
solving it used disinfectant such as ultraviolet and antibiotic. Using antibbiotic was effective to solve the
problem but it had a bad effect to human and environment health. In addtion that used antibiotic will
disturb the dinamic of biological balance of microorganism or capability of larvae to adapt with
environment (Maeda et al., 1992; Haryanti, et al., 1999; Susanto et al 2005). Other reason said that
presently the awwarnes of consumer to food safety was high. Hence, the better solution for improving of
rearing technic is by environment friendly approach with using probiotic bacteria.
The paper was presented in Indonesian Aquaculture 2009, Manado Nort Sulaawesi Indonesia
Center for Brackhiswater Aquaculture Development Takalar, Indonesian Crab Development Center, Marine Affairs and Fishery Department of Indonesia
On the rearing of swimming crab larvae, reducing of water quality was commonly caused by
accumulation of organic mater from metabolism and feed watste (Effendy et al., 2005a). High organic
mater dissolved in water media giving the chance to bacteria for growth and developed due to
population dinamic of bacteria depended to supply nutrition (Liu & Han, 2004). Developing of
decomposser bacteria mainly negative bacteria has potentially infect the crab larvae. This infection
caused larva death if its has a lowwer immunity and bacterial density were more than 103 CFU/ml.
(Lavilla-Pitogo et al., 1998 in Effendy et al 2006a).
The preventive effort of bacterial infection was developed, including probiotic application for
reducing the growth of opportunistic bacteria such as Vibrio sp in water media. Probiotic was proven to
be able to reduce the growth of Vibrio sp. until 102 CFU/ml (Effendy et al;, 2006a), however probiotic
bacteria has a short life time so needed substrate which can provide the feed. Molases contains high
sugar (about 45-58%) including glucosa, sucrosa, and fructosa (Paturau 1982) (so its can become a
substrate and feed source for growth of probiotic bacterial. Contrary addition of molases withought
bacteria probiotic will invite pathogen bacteria.
Recenly, application of probiotic and molases was more commonly used for bacterial culture in
laboratoroum scale and shrimp pond. but effectiviness of application of probiotic and molases in rearing
water media of swimming crab larvae still unverified, hence It should be conducted the research more
for getting the effective application method.
This study was aimed to know the effectiveness of probiotic bacteria and molases toward
incerasing the survival rate of swimming crab larvae.
MATERIALS AND METHODS
Probiotic and Molases
Probiotic bacteria used from comercial product “produced from Develop TM” and molases from
the sugarcan factory in Takalar – South Sulawesi Indonesia.
Rearing of larvae
Swimming crab larvae (Portunus pelagicus) of zoeae stage was stocked in to fiber pond 250 L
with stocking density 60 ind/ml. Larvae has reared from zoeae-1 to Crablet-1 for 15 days. Larvae zoeae-
1 to 4 was fed by rotifer Branchionus plicatilis with density 10-15 ind/ml and its enriched with fatty acid
Ω3 – HUFA and multivitamin with concentration 100 ppm for each suplement, whereas Artemia salina
was fed to larvae of zoeae-3 with density 0,5- 3 ind/ml. Feeding was carried out for every day.
This study was conducted with two treatments (A) swimming crab larvae was reared using
probiotic (B) reared using probiotic and molases. Probiotic and molases was introduced into the water
media from zoeae-1 and then given every 3 day. Probiotic powder was dissolved into water about 10
ppm, and then it introduced to the water media of larvae after water exchange. Whereas molases
introduced about 100 ppm into 10 ppm probiotic solution, then aerated for 6 hours prior to introduce to
rearing water media. Water exchange was conducted conterminoused with siphon activities.
1. Survival Rate
Survival rate of swimming crab larvae was measured for each growth stage. Survival rate is the
procentage of number of larvae at the initial and end rearing. The formula of survival rate based on
Effendie (1979) as follow :
S = x 100%
Dimana: S = Survival rate (%)
Nt = Population at the end of rearing
No = Population at the initial of rearing
2. Growth Stage of Larvae
The development of larvae was observed with collected the larvae samples of each stage about
30 larvaes under microscope. The observed comprised lenght of carapace, body weight, and larvae
development for each stage (Zoeae-1 to crablet-1).
3. Population of Bacteria
Computing of population of bacteria were to calculate the Bacteria total and Vibrio sp total.
Bacteria total was conducted to know the population bacteria total. Calculation method was used culture
media of natrium agar (NA). Wherease calculation the Vibrio total used TCBSA culture (thiosulfat citrate
bilesalt sucrose agar),
4. Water Quality
Parameter of water quality was observed every day such as Dissolved Oxygen (DO), pH,
Temperature, salinity, ammonia and nitrit. Water sampels were carried out prior to water exchange
mainly for pH, ammonia and nitrit, whereas for DO, temperature, and salinity were directly carried out in
pond using portable DO YSI 55, for measuring DO and temperature and repractometer Yamato for
measuring salinity. Water exchange was conducted after 3th day for 30% of volume and then increased
until 80% of volume while entered to megalop stage or followed the water quality of water media.
RESULTS AND DISCUSSION
Results showed that treatment of probiotic affected to growth stage from zoeae (zoeae 4) to
megalops stage on 10th day (with 10% megalops) and mogealops to crablet on 15 th day (10% crablets).
Whereas treatment by probiotic and molases affected to growth stage from zoeae (zoeae 4) to
megalops stage on 9th day (with 90% megalops) and mogalops to crablet on 14 th day (20% crablets).
With that results showed that addition of molases was accelerated growth stage from zoeae to megalops
and megalops to crablet (Table 1 and Figure 1). Becouse larvae growth were more normal withought
Table 1. Growth stage development of swimming crab larvae in rearing periode
PERIOD ROBIOTIC PROBIOTIC AND MOLASES
1 Zoeae-1 Zoeae-1
2 Zoeae-1 Zoeae-1
3 Zoeae-1 Zoeae-1(10%)/ Zoeae-2 (90%)
4 Zoeae-2 Zoeae-2
5 Zoeae-2 Zoeae-3
6 Zoeae-3 Zoeae-3
7 Zoeae-3 Zoeae-4
8 Zoeae-4 Zoeae-4
9 Zoeae-4 Zoeae-4 (40%)/Megalop-1 (60%)
10 Zoeae-4(90%)/ Megalop-1 (10%) Megalop-1
11 Megalop-1 Megalop-2
12 Megalop-2 Megalop-3
13 Megalop-3 Megalop-4
14 Megalop-4 Megalop-5 (80%)/Crablet-1(20%)
15 Megalop-5/Crablet-1 (10%) Crablet-1
Treatment of probiotic bacteria showed that carapaces were growth from 0,16 to 0,57 mm and
weight from 0,00004 to 0,00059 g for 10 days rearing of zoeae stage and in the stage megalops
carapaces were growth from 0,57 to 1,14 mm and weight from 0,00050 to 0,00117 g for 5 day rearing of
megalops. For treatment probiotic and molases showed that carapace was growth from 0,14 to 0,53
mm and weight from 0,00006 to 0,00043 g for 9 days rearing of zoeae stage and in the stage megalops
carapaces were growth from 0,53 to 1,18 mm and weight from 0,00043 to 0,00113 g for 5 days rearing
of megalops and day of 15th all megalops were entered to crablet stage (Figure 2).
The interesting evidances in treatment of probiotic and molases on 3th day, 90% of zoeae-1 were
entered to zoeae-2, also in 9th day, 60% of zoeae-4 was entered to Megalops-1. Finally in 14th day 60%
of megalops were entered to crablet and in 15 th all megalops were entered to crablet and this fact was
not find in treatment probiotic only (Figure 2). In 15th day it results showed lenght of carapace and
weight body of treatment probiotic and molases were more lenght and weight than treatment probiotic
only but not significantly different (P>0,05). We conclude that addition of molases into water rearing
media of swimming crab tends to reduce the growth stage (zoeae to crablet stage) and tend more
weight and lenght, as larvae growth was optimal. But unsignificant data between both treatment might
be caused by molases and C-N ratio of the feed were not optimal, despite the Vibrio sp population was
succesfully controlled (Table 2).
Population of Growth stage (%)
1 2 3 4 5 6 7 8 9 10 11 12 13 14 15
100 Rearing of the day
Population of Growth stage (%)
Zoeae-1 Zoeae-2 Zoeae-3 Zoeae-4 Megalops Crablet
1 2 3 4 5 6 7 8 9 10 11 12 13 14 15
Zoeae-1 Zoeae-2 Zoeae-3 Zoeae-4 Megalops Crablet
Figure 1. Growth stage development of swimming crab larvae (A) Treatment of Probiotic and molases
and (B) Treatment of probiotic only.
1 3 5 7 9 11 13 15
Probiotic Rearing period
Lenght of carapace (mm)
1 3 5 7 9 11 13 15
Figure 2. Growth of swimming carb larvae in rearing period (A) weight body and (B) lenght of carapace
The survival rate was significantly different (P<0.05) for stage zoeae-3,4 and megalops (Figure 3),
but not significatly different for stage of zoeae-2 and crablet-1 (P>0,05). Treatment of probiotic showed
that survival rate are 100%, 73,33%, 55%, 35%, 21,67% and 15% for zoeae-1, zoeae-2 zoeae-3, zoeae-
4, mengalops and crablet-1 resvectively. Those results are more less than treatment of probiotic and
molases, namely 100%; 85%; 78,33%; 55%; 40%; and 26,67 % resvectivelly (Figur 3). Thereby
Probiotic and molases played an important role in increasing survival rate of swimming crab larvae.
Survival rate (%) Probiotic and Molace
Z1 Z2 Z3 Z4 M C
Figure 3. Survival rate of each stage on rearing period of swimming crab larvae
Population of Bacteria
The investigation results of bacteria total and Vibrio sp showed that treatment of probiotic
bacteria, both population on stage zoeae-1 were 2,4 X 103 cfu/ml and 2,0 X 102 cfu/ml and increased at
the end of rearing period with population density about 6,0 X 104 cfu/ml and 4,2 X 103 cfu/ml
resvectivelly. Whereas on the treatment of probiotic and molases showed that both population density
were 2,0 X 104 cfu/ml and 3,2 X 103 cfu/ml and also increased at the end of rearing period with
population density about 4,2 X 106 cfu/ml and 2,0 X 102 cfu/ml resvectivelly (Table 2).
Table 2. Population density of bacteria total dan Vibrio sp. on each growth stage (cfu/ml).
Treatments Parameters Growth Stage
Zoeae-1 Zoeae-2 Zoeae-3 Zoeae-4 Megalop
Probiotic Bacteria Total 2,4 X 103 2,0 X 104 3,2 X 104 1,1 X 103 6,0 X 104
Vibrio sp 2,0 X 102 2,0 X 103 1,2 X 103 2,1 X 102 4,2 X 103
Probiotic and Bacteria Total 2,0 X 104 3,4 X 104 4,6 X 104 4,2 X 106 4,2 X 106
Molases Vibrio sp 3,2 X 103 3 X 102 3,2 X 102 2,2 X 102 2 X 102
Base on the above (Table 2) indicated that addition probiotic and molases into water media could
reduce the population density of Vibrio sp until 102 cfu/ml. This evidence appeared on all growth stage,
except in zoeae-1 due to adaptation process, growth process of probiotic bacteria, and has a high
bacteria total density. Both treatments also had the different population density. Treatment of probiotic
and molases was higher population density of bacteria total than treatment of probiotic only. Its indicated
that molases plays and important role in providing nutrition for probiotic.
Acceleration of growth of probiotic bacteria after introduction molases was high, so its won the
conpetition with other bacteria including pathogen bacteria such as Vibrio sp. In addition, probiotic
bacteria released the antibiotic or antimicrobial. In the other hand probiotic bacteria become inhibitor for
other bacteria (Irianto, 2005) or controlled pathogen bacteria. Life cycle of probiotic in both treatments
was different which indicated by population of bacteria total in molases treatment was higher and
population of Vibrio sp was controlled in 102 cfu/ml. Thereby using probiotic will have the benefit if
rearing water media has enough feed for bacteria.
Comparison in the both treatment showed that the presence of population of probiotic bacteria will
be able to inhibit the population of bactera pathogen. Aplication probiotic in the water media will have
benefit more if probiotic could growth and develop well, furthermore nutrition should be provided. Used
molases or mixture of rice and bran was enough for supplying nutrition of probiotic which its had fuction
as source of organic carbon (Suprapto, 2006). Also that contains the good ratio of carbon-nitrogen will
more support the growth of probiotic (Goldman & Dennett 2000) also larvae. Survival factor of probiotic
in inhibiting growth rate of bacteria patogen was important (http://horizonpress.com/hsp/pro.html), so
addition of molases as substrat and nutrition also important.
Water quality monitoring for rearing period appeared that in the both treatment were similarly
(Table 3) and in the rang of good water quality for rearing of swimming crab larvae (Effendy et al , 2005;
Nurcahyono & Kasturi, 2008). Therefore water quality were not significat effect to the both different
result. However using of both treatment was reduced the posibillity of dropped water quality. It was clear
that addition of bacteria probiotic and molases controlled water quality and affected to growth stage
Table 3. Water quality performance in both treatment probiotic and probiotic with molases of rearing
swimming crab larvae (Portunus pelagicus,linn)
Probiotic Probiotic dan Molases
Dissolved Oxygen (mg/l) 4,8 – 5,6 4,6 – 6,2
pH 7,8 – 8,6 8,1 – 8,4
Temperature (0C) 27 – 31 27 – 31
Salinity (ppt) 33 – 35 33 -35
Ammonia (mg/l) Not detected Not detected
Nitrit (mg/l) Not detected Not detected
CONCLUSION AND RECOMENDATION
Application of the probiotic bacteria with molases in water media of swimming crab larvae
(Portunus pelagicus,linn) tends to increase the survival rate, body weight, lenght of carapace, and able
to accelerate the growth of zoeae stage, in addition also controlled the population growth of Vibrio sp
(102 cfu/ml). Furthermore using probiotic with molases was effective in improving of rearing water media
on mass producing crablet of swimming crab.
For the furter it should be conducted the detail study in using of concentration molases, probiotic
density and other combination substrat for getting the high survival rate and growth such as C-N ratio.
Effendy, S., Faidar, Sudirman, E Nurcahyono., 2005a. Perbaikan Teknik Pemeliharaan Larva Pada Produksi
Masal Benih Rajungan Portunus pelagicus. Prosiding Pertemuan Teknis Lintas UPT Pusat Budidaya Air
Payau dan Laut tanggal 19 – 21 Juli 2005 di Imperial Aryaduta Makassar.
Effendy, S., Faidar, Sudirman, E Nurcahyono., 2005b. Perkembangan Larva Rajungan Portunus pelagicus Pada
Produksi Masal Pascaintroduksi Artemia salina dan Brachionus plicatilis diperkaya Asam Lemak.
Prosiding Konferensi Akuakultur Nasional di Sahid Jaya makassar, tanggal 23 – 25 November 2005.
Effendy, S., Sudirman, Faidar. dan E Nurcahyono., 2006. Efektivitas Bakteri Probiotik pada Produksi Massal
Larva Rajungan Portunus pelagicus Linn. Makalah Dipresentasikan pada Konferensi Nasional
Masyarakata Akuakultur (MAI) di Surabaya.
Effendy, S., Sudirman, Bahri, S., E Nurcahyono., Batubara, H. dan Syaichudin, M., 2006b. Petunjuk Teknis
Pembenihan Rajungan (Portunus Pelagicus Linnaeus). Balai Budidaya Air Payau Takalar.
Goldman, jC Dennett MR. 2000. Growth of marine bacteria in the batch and continuous cultur under carbon and
nitrogen limitation. Limnol Oceanogr 45(4):789-800.
Irianto, A., 2005. Budidaya Perikanan Berkelanjutan Melalui Pemanfaatan Probiotik. InfoMai Vol. 4 : 1
Juwana, S., 1997. Tinjauan Tentang Perkembangan Penelitian Budidaya Rajungan (Portunus pelagicus) Volume
XXII. Pusat Penelitian dan Pengembangan Oseanologi – LIPI, Jakarta.
Liu F and Han W. 2004. Reuse strategy of wastewater in prawn nursery by microbial remediation aquacullture.
Nurcahyono, E., Kasturi., Makkulau Sultan. 2008. Manajemen Pembehihan Rajungan Dalam Upaya Peningkatan
Produksi Rajungan (Portunus Pelagicus,Linn). Makalah Dipresentasikan pada Indonesian Aquaculture di
Paturau, JM. 1982. By-Product of The Cane Suger Industry. Amsterdam: Elselvier Scientific Publishing Company.
Sumpton, W., Gaddes, S., Mc Lennan, M., Campbell, M., Tonks, M., Good, N., Hagedoorn, W., Skilleter, G.,
2003. Fisheries Biology and Assessment of the Blue Swimmer Crab (Portunus pelagicus) in
Queensland. The State of Queensland, Dept. of Primary Industries, and the Fisheries Research and
Susanto,B., I Setyadi, D Syahidah,M Marzuqi, I Rusdi.2005. Penggunaan Bakteri Probiotik Sebagai Kontrol
Biologi DalamProduksi Massal Benih Rajungan (Portunus pelagicus). Jurnal Penelitian Perikanan
Indonesia 11 (1) 15 - 23.