Abstract
Objective: The present study reported the isolation of lactic acid bacteria from the shrimp gastrointestinal tract and evaluation of its antagonistic activity against pathogenic bacteria. Methods/ statistical analysis: Lactic bacteria from the gastrointestinal tract of alive healthy shrimp were screened on sterilized MRS medium. Vibrio sp. inhibition by isolates was conducted by the agar-well diffusion assay. The isolates were identified by 16S rRNA sequencing. Antibacterial activity was investigated against 25 Vibrio sp. isolates, Staphylococcus aureus ATCC 25023, and Escherichia coli ATCC 85922. Hemolytic activity, salt, and antibiotic resistance were also evaluated. Findings: Total of 22 isolates exhibited Vibrio parahaemolyticus inhibition with antagonistic activities ranged from 220 AU/ml to 460 AU/ml. Sequence analysis of 16S rRNA indicated the 5 isolates belong to Weissella cibaria species, namely HN02, HN03, HN04, and HN06 AND HN07. Among these, W. cibaria HN05 had highest antagonistic activity and exhibited the broad inhibition spectrum against Vibrio sp. as well as E. coli ATCC 85922 but not S. aureus ATCC 25023. β-hemolytic and salt resistance assays showed that W. cibaria HN05 had no β-hemolytic activity and grew well in medium containing 5% of salt. Antibiotic resistance analysis indicated W. cibaria HN05 was suppressed by ampicillin, chloramphenicol, oxytetracycline hydrochloride, and chlortetracycline hydrochloride. Application/improvements: This is the first study on isolation and evaluation pathogenic bacteria including Vibrio sp. inhibition by W. cibaria, which is isolated from shrimp gastrointestinal tract. Keywords: Antagonistic Activity, Gastrointestinal Tract, Shirmp, Weissella cibaria, Vibrio sp.
Highlights
Pacific white shrimp, Litopenaeus vannamei, is the most popular shrimp species due to their survival advantages such as fast growth rate and strong adaptability to the environment [1]
Isolates displayed as Grampositive bacteria and had the V. parahaemolyticus strong inhibition were identified by the 16S RNA fragment sequencing method
DNA sequence alignment confirmed 5 isolates (HN02, HN03, HN04, HN05, and HN07) have 100% similarity to Weissella cibaria species and they were deposited on Genbank database as accession number of MK989999, MK990000, MK990001, MK990002, and MK990004, respectively
Summary
Litopenaeus vannamei, is the most popular shrimp species due to their survival advantages such as fast growth rate and strong adaptability to the environment [1]. In 2009, acute hepatopancreatic necrosis disease (AHPND) was first reported in China disease outbreaks occurred in various countries in Southeast Asia and America, resulting in up to 100% mortality. Vibrio species such as V. parahaemolyticus and V. harveyi were demonstrated as causative agent of AHPND [3,4]. Response for disease on shrimp including luminous vibriosis, tail necrosis, shell disease, red disease, loose shell syndrome, white gut disease, white spot, etc [5,6,7] It is estimated shrimp farming industry lost more than 1$ billion US dollar per year [8]. Developing an appropriate strategy to control Vibrio species in aquaculture increases the productivity and enhance the product safety
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