Effect of a Probiotic and Herbal

Additives on Growth, Survival and

Disease Resistance of Striped Murrel
M.A. Haniffa, B. Kavitha Bharathi, I. Viji Margaret and Bilal Ahmad Paray

A mong air-breathing In the last few years probiotics and herbs have residues of antibiotics that
freshwater food fishes, become an integral part of culture prevention ultimately diffuse into the
snakeheads (commonly called for promoting growth and disease resistance. aquatic environment (Ellis
murrels in India) constitute the This strategy offers many advantages to 1991, Smith et al. 1994).
most common and dominant overcome the limitations and side effects of Application of modern tools is
group. They fetch very high antibiotics and other drugs and also leads to necessary to increase disease
prices because the flesh quality high production through enhanced growth resistance and promote murrel
is high in terms of taste and and disease preventio. In aquaculture, the range aquaculture (Ghosh et al.
texture, with a low fat content, of probiotics evaluated for use is considerably 2002).
fewer intramuscular bones, wider than in terrestrial agriculture. An alternative and safer
and perceived medicinal value way to tackle Aeromonas
(Haniffa et al. 2004). The striped murrel Channa striata (Fig. infection is an eco-friendly development of resistance by the target
1) is a promising species for aquaculture in Asian countries but species through application of probiotics (Ringo and Birkbeck
is constrained by the availability of seed and feed. Murrels are 1999, Irianto and Austin 2002) and herbs (Guojun et al. 2008).
carnivorous, piscivorous and cannibalistic in nature. These immunostimulants enhance the humoral and cellular
Murrels are affected by Epizootic Ulcerative Syndrome immune responses in specific and non-specific ways, thereby
(EUS), primarily caused by the fungus Aphanomyces invadans reducing the risk of diseases.
and secondarily by the bacterium Aeromonas hydrophila In the last few years probiotics and herbs have become
(Karunasagar et al. 1995). Infection by A. hydrophila is generally an integral part of culture prevention for promoting growth
associated with poor environmental conditions (Grabowshi et al. and disease resistance. This strategy offers many advantages
2004). Invasion by the pathogen results in skin lesions that lead to overcome the limitations and side effects of antibiotics and
to high mortality. Common control measures of the diseases rely other drugs and also leads to high production through enhanced
on antibiotics, chemotherapeutants and vaccines. However, using growth and disease prevention (Das et al. 2008, Sahu et al.
antibiotics for treatment is problematic, relating to the acquisition 2008). In aquaculture, the range of probiotics evaluated for use is
of genes that result in antibiotic-resistant strains of bacteria and considerably wider than in terrestrial agriculture.

FIGURE 1. C. striatus affected by the disease Epizootic Ulcerative Syndrome. FIGURE 2. Feed additives and semi-moist feed.

64 J U N E 2 013 • W O R L D AQ UACU LT U R E • W W W.WA S .O R G

 TOP LEFT, FIGURE 3. Injection of A. hydrophila to C. striatus fingerlings.
TOP RIGHT, FIGURE 4. Effect of feed additives on weight gain in C. striatus
fingerlines fedd different feeds.
LEFT, FIGURE 5. Fingerlings of C. striatus showing skin lesions challenged by
A. hydrophila.

The goal of the investigation described here was to assess

the effects of a common probiotic bacterium, Bacillus subtilis,
and herbal additives Indian almond Terminalia catappa and
aloe Aloe vera on growth performance, survival and disease
resistance of striped murrel fingerlings challenged by A.
hydrophila.

Effects on Growth Rate

Four experimental diets were prepared (Table 1). Each of
three diets contained one of the following feed additives (Fig.
2): Bacillus subtilis, Indian almond Terminalia catappa and
TABLE 1. Semi-moist feed for Channa striata. aloe Aloe vera. A fourth (control) diet was prepared without
feed additives. Ingredients were mixed with feed additives and
Ingredient % autoclaved, semi-moist feeds were prepared and stored at -20 C.
Striped murrel fingerlings (1.0±0.2 g/fish) were obtained
Fishmeal, anchovy 26.9 from the Centre for Aquaculture Research and Extension
Soybean flour 25 (CARE) Aquafarm, St. Xavier’s College, Palayamkottai, Tamil
Jawala, Acetes sp. 20 Nadu, India and stocked into cement tanks (300 L) filled with
Tapioca meal 10.9
groundwater at 10 fingerlings per tank in triplicate for each
treatment. Fingerlings were fed 5 percent of body weight daily,
Wheat flour 10
divided into three feedings, for 50 days and the feeding rate
Sunflower oil 5.8 was adjusted every 10 days. Growth parameters were calculated
Monosodium phosphate 0.3 using standard methods (Adewolu 2008).
Mineral premix 0.5 Growth of striped murrel fed diets with a probiotic
Aqua Saver 0.3 bacterium and aloe were not different from striped murrel fed
Vitamin and Mineral premix 0.1
the control diet. However, growth of striped murrel fed the diet
with Indian almond was substantially greater than that of fish
Feed additive 0.2
fed the control diet (Table 2). Some herbs promote cellular lipid
a Vitamin mixture providing the following concentration per kilogram
and fatty acid utilization and protein accumulation resulting in
diet; vitamin A 5000 IU; vit D 400 IU; vit E 20 mg; thiamin
good growth performance (Ji et al. 2007). The growth increase
in rohu Labeo rohita fed with herbal supplemented diet was a
mononitrate (B1) 4 mg; riboflavin (B2) 6 mg; nicotinamide 50
mg; pyridoxine hydrochloride 3 mg; calcium pentohenate 10 mg;
cyanocobalamine (B12) 2mg; ascorbic acid (vit C) 100 mg; biotin 0.1
result of improved food utilization and high protein synthesis
mg. b Trace mineral mix use providing the following concentration (ppm) (Johnson and Banerji 2007).
copper 10; iron 100; manganese 50; zinc 50; cobalt 0.05; and iodine
0.1 (Dhanaraj and Haniffa 2011). Feed additives- B. subtilis, T. Effects of Disease Challenge
catappa, A. vera. A. hydrophila was isolated from diseased striped murrel
(CONTINUED ON PAGE 66)

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 TABLE 2. Growth Performance of C. striatus fingerlings fed on different diets.

Parameters Aloe vera Bacillus subtilis Terminalia catappa Control

Initial length (cm) 4.6±0.4 4.7±0.3 4.9±0.4 4.9±0.4

Final length (cm) 7.9±0.5 8.2±0.8 9.7±0.4 7.9±0.5
Initial weight (g) 1.0±0.2 1.0±0.2 1.0±0.2 1.0±0.2
Final weight (g) 3.9±0.7 3.9±0.7 7.3±0.6 3.9±0.5
Weight gain (g) 2.9 2.9 6.3 2.9
SGR (%/d) 2.7 2.9 4.0 2.6
Condition Factor 0.79 0.77 0.80 0.79
Survival (%) 100 90 90 60

TABLE 3. Mortality of C. striata fingerlings challenged by A. hydrophila.

Diet Hours post-challenge

24 48 72 96 120 144 168 Total

Control * * * 1/10 * 1/9 2/8 4/10
Terminalia catappa * * * * 1/10 * * 1/10
Aloe vera * * * * * * * 0/10
Bacillus subtilis * * * * * 1/10 * 1/10

66 J U N E 2 013 • W O R L D AQ UACU LT U R E • W W W.WA S .O R G

collected from the wild. A subculture was maintained on tryptone Grabowshi, L.D., S.E. LaPatra and K.D. Cain. 2004. Systemic
soy agar slopes (Himedia) at 4 C and was routinely tested for and mucosal antibody response in tilapia, Oreochromis
pathogenicity. A stock culture in tryptone soy broth was stored at niloticus (L.), following immunization with Flavobacterium
-20 C to provide stable inoculum throughout the study (Dhanaraj columnare. Journal of Fish Diseases, 27:573-581.
and Haniffa 2011). After the 50-d feeding trial, experimental fish Guojun, Yin, Ardo Laszlo, Jeney Zsigmond, Xu Pao and
were injected intraperitoneally (Fig. 3) with one-day-old culture Jeney Galina. 2008. Chinese herbs (Lonicera japonica and
of A. hydrophila (100 µL/fish) at 106 cfu/mL. All fish were Ganoderma incidum) enhance non-specific immune response
observed daily for one week to record clinical signs and mortality. of tilapia, Oreochromis niloticus and protection against
Survival of striped murrel fed diets supplemented with Aeromonas hydrophila. Deseases in Asian Aquaculture VI:
aloe was greatest (Table 3). Survival of striped murrel fed diets 269-282.
supplemented with Bacillus and Indian almond was greater than Haniffa, M.A., K Marimuthu., M. Nagarajan., A.J. Arokiaraj
that of fish fed the control diet. The improved survival suggests and Kumar, D. 2004. Breeding behaviour and parental care
that production of metabolites by probiotics suppressed the of the induced bred spotted murrel Channa punctatus under
pathogenicity of A. hydrophila (Fig. 5) and ultimately resulted captivity. Current Science 86:1375-1376.
in better survival and growth of the host. Aloe was effective as Haniffa, M.A and Abdul Kader Mydeen. 2011. Hematological
a disease suppressant and antibacterial agent in striped murrel. changes in Channa striatus Experimentally infected by
Substances in aloe are known to have anti-bacterial and anti- Aeromonas hydrophila. Bioresearch Bulletin 4:246-253.
fungal effects (Klein and Penneys 1988, Kim et al. 1999, Sunitha Irianto, A and Austin B. 2002. Use of probiotics to control
2012). Aloe is recommended as a feed supplement for treatment furunculosis in rainbow trout, Oncorhynchus mykiss
of EUS. (Walbaum). Journal of Fish Diseases 25:333-342.
Ji, S.C., O. Takaoka, G.S. Jeong, S.W. Lee, K. Ishimaru, M. Seoka
and K. Takii. 2007. Dietary medicinal herbs improve growth
Notes and some nonspecific immunity of red seabream Pagrus
M.A. Haniffa, Centre for Aquaculture Research and Extension major. Fisheries Science 73:63-69.
(CARE), St. Xavier’s College, Palayamkottai – 627 002, Tamil Johnson, C. and A. Banerji. 2007. Influence of extract isolated
Nadu, India. Email: [email protected], Website : www. from the plant Sesuvium portulacastrum on growth and
caresxc.org metabolism in freshwater teleost, Labeo rohita (Rohu). Fishery
Technology 44(2):229-234.
Karunasagar, I., G. Sugumar and I. Karunasagar. 1995. Virulence
Acknowledgments characteristics of Aeromonas sp. isolated from EUS-affected
Authors acknowledge the financial assistance received from fish. Pages 307S–314 In: M. Shariff, J. R. Arthur and R. P.
ICAR-NAIP Sub Project entitled, “A Value Chain on Murrel Subasinghe, editors. Diseases in Asian Aquaculture Vol. II.
Production in Tamilnadu and Orissa” (F.No.1(5)/2007-NAIP dt. Manila, Philippines: Fish Health Section, Asian Fisheries
22 Aug. 2008). We are grateful to Rev. Dr. Joseph S.J., Principal Society,.
and Consortium Leader, St. Xavier’s College (Autonomous), Kim, H.S., S. Kacew and B.M. Lee. 1999. In vitro
Palayamkottai, for providing the necessary facilities. chemopreventive effects of plant polysaccharides (Aloe
barbadensis Miller, Lentinus edodes, Ganoderma lucidum
and Coriolus versicolor). Carcinogenesis 20:1637-1640.
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