J. Life Sci. Biomed. 6(1): 10-14, Jan 30, 2016
Life Science and Biomedicine
Antimicrobial Activity of Bacillus cereus and Bacillus thuringiensis on
Pathogenic Vibrio harveyi in Litopenaeus vannamei
Marissa Maya Masitoh1 , A. M. Hariati2 and M. Fadjar2
1Master Program on Faculty of Fishery and Marine Science, University of Brawijaya, Malang 65145, Indonesia
2Department of Aquaculture, University of Brawijaya, Malang 65145, Indonesia
ABSTRACT: The bacterial strain of Bacillus cereus and Bacillus thuringiensis has been known to produce
antimicrobial activity against pathogenic Vibrio harveyi. The effect of B. cereus and B. thuringiensis were tested
by in vitro and in vivo. In vitro test was used to analyze antagonism characteristic of bacteria using the paper
disc diffusion method. In vivo test was applied to evaluate antimicrobial activity of B. cereus and B.
thuringiensis (105 CFU ml-1) on survival rate and histopathology of Litopenaeus vannamei challenged with V.
harveyi. The results showed that B. thuringiensis had a greater inhibitory activity of 18.60–35.97 mm. Both
Bacillus bacteria treatment resulted in survival rate of 100%, compared with 75% in the treatment without
Bacillus. It can be concluded that B. cereus and B. thuringiensis have potential applications for controlling
pathogenic V. harveyi in shrimp aquaculture.
Key words: Bacillus cereus, Bacillus thuringiensis, Inhibitory Activity, V. harveyi.
Aquaculture is the world’s fastest growing food production sector. However, disease outbreaks have caused
serious economic losses in several countries. Vibrio species are among the most important bacterial pathogens of
cultured shrimp. They are responsible for several diseases and mortalities up to 100% due to vibriosis have been
reported . Using antibiotics in potential negative consequences of using antibiotics in aquaculture for the
prophylactic treatment of diseases are the development of drug resistant bacteria and reduced efﬁcacy of
antibiotic treatment for human and animal diseases .
In the search for more effective and environmentally friendly treatments, using bacteria like Bacillus
provides a solution to these problems. Bacillus have antimicrobial (bacteriocin) which usually occurs in all
growth phases and finishes at the end of logarithmic phase . Luis-Villaseñor et al.  investigated that the
effect of Bacillus showed trait inhibitory to Vibrio and ability to adhere and grow on intestinal mucus.
The purpose of this study was to investigate antimicrobial activity of B. cereus and B. thuringiensis against
Vibrio harveyi under in vitro and in vivo conditions.
MATERIAL AND METHODS
A virulent strain of V. harveyi, was used as a pathogenic strain. Strain were taken from the stock culture
collection of our laboratory. B. cereus and B. thuringiensis, obtained from isolated from the gastrointestinal of L.
vannamei and cultured in duplicate in the general media (nutrient agar with 1.5% w/v NaCl) for 18-24 hours at a
temperature of 30oC. Pure isolates were taken after subculture on Tryptic Soya Agar (TSA) was used as an
The methods of paper disc diffusion assays were used in this study. Both groups of the bacterial strains (the
tested strains including the compared strain and the pathogenic strain) were briefly grown in tryptic soya broth
(TSB), incubated at 32oC for 24 h. After 24 h, each sterilised paper disc was immersion with B. cereus and B.
thuringiensis with different concentration (104, 105 and 106 CFU ml-1). And then placed on the surface of an agar
plate (TSA) which was previously inoculated with the indicator pathogen at a concentration of about 107 CFU
ml-1. The plate was then incubated at 32o C for 24 h and the inhibition zone around paper disc was recorded.
To cite this paper: Masitoh M. M., Hariati A. M. and Fadjar M. 2016. Antimicrobial Activity of Bacillus cereus and Bacillus thuringiensis on Pathogenic Vibrio harveyi
in Litopenaeus vannamei. J. Life Sci. Biomed. 6(1): 10-14.