MATERIALS AND METHODS: A. hydrophila and E. tarda were isolated using glutamate starch phenol red and xylose lysine deoxycholate (Merck, Germany) as a selective medium, respectively. All the suspected bacterial colonies were identified using conventional biochemical tests and commercial identification kit (BBL Crystal, USA). Susceptibility testing of present bacterial isolates to 16 types of antibiotics (nalidixic acid, oxolinic acid, compound sulfonamides, doxycycline, tetracycline, novobiocin, chloramphenicol, kanamycin, sulfamethoxazole, flumequine, erythromycin, ampicillin, spiramycin, oxytetracycline, amoxicillin, and fosfomycin) and four types of heavy metals (mercury, chromium, copper, and zinc) were carried out using disk diffusion and two-fold agar dilution method, respectively.
RESULTS: Three hundred isolates of A. hydrophila and E. tarda were successfully identified by biochemical tests. Antibiotic susceptibility testing results showed that 42.2% of the bacterial isolates were sensitive to compound sulfonamides, sulfamethoxazole, flumequine, oxytetracycline, doxycycline, and oxolinic acid. On the other hand, 41.6% of these isolates were resistant to novobiocin, ampicillin, spiramycin, and chloramphenicol, which resulted for multiple antibiotic resistance index values 0.416. Among tested heavy metals, bacterial isolates exhibited resistant pattern of Zn(2+) > Cr(6+) > Cu(2+) > Hg(2+).
CONCLUSION: Results from this study indicated that A. hydrophila and E. tarda isolated from coinfected farmed red hybrid tilapia were multi-resistant to antibiotics and heavy metals. These resistant profiles could be useful information to fish farmers to avoid unnecessary use of antimicrobial products in the health management of farmed red hybrid tilapia.
METHODS AND RESULTS: The crude extracts of E. pubescens were obtained through methanol extraction, and evaluated for antimicrobial activities. From this extract, 1,7-bis(3,4-dihydroxyphenyl)heptan-3-yl acetate (etlingerin) was isolated. When compared to curcumin (a compound with a similar chemical structure), etlingerin showed twofold lower minimum inhibitory concentration values while also being bactericidal. Through time kill assay, etlingerin showed rapid killing effects (as fast as 60 min) against the Gram-positive bacteria (Staphylococcus aureus ATCC 43300 and Bacillus subtilis ATCC 8188). Further assessment revealed that etlingerin caused leakage of intracellular materials, therefore suggesting alteration in membrane permeability as its antimicrobial mechanism. Cytotoxicity study demonstrated that etlingerin exhibited approximately 5- to 12-fold higher IC50 values against several cell lines, as compared to curcumin.
CONCLUSIONS: Etlingerin isolated from E. pubescens showed better antibacterial and cytotoxic activities when compared to curcumin. Etlingerin could be safe for human use, though further cytotoxicity study using animal models is needed.
SIGNIFICANCE AND IMPACT OF THE STUDY: Etlingerin has a potential to be used in treating bacterial infections due to its good antimicrobial activity, while having potentially low cytotoxicity.
METHODS AND RESULTS: The pulp of red pitahaya and the leaves of red spinach were extracted using methanol followed by subfractionation to obtain betacyanin fraction. The anti-biofilm activity was examined using broth microdilution assay on polystyrene surfaces and expressed as minimum biofilm inhibitory concentration (MBIC). The betacyanin fraction from red spinach showed better anti-biofilm activity (MBIC: 0·313-1·25 mg ml-1 ) against five Staph. aureus strains while the betacyanin fraction from red pitahaya showed better anti-biofilm activity (MBIC: 0·313-0·625 mg ml-1 ) against four P. aeruginosa strains. Both betacyanin fraction significantly reduced hydrophobicity of Staph. aureus and P. aeruginosa strains. Numbers of Staph. aureus and P. aeruginosa attached to polystyrene were also reduced without affecting their cell viability.
CONCLUSION: Betacyanins can act as anti-biofilm agents against the initial step of biofilm formation, particularly on a hydrophobic surface like polystyrene.
SIGNIFICANCE AND IMPACT OF THE STUDY: This study is the first to investigate the use of betacyanin as a biofilm inhibitory agent. Betacyanin could potentially be used to reduce the risk of biofilm-associated infections.