Purification and characterization of bioactive compounds extracted from Suaeda maritima leaf and its impact on pathogenicity of Pseudomonas aeruginosa in Catla catla fingerlings

Abstract

Incidence of various dreadful microbial infections and the development of antibiotic resistance by infection causative microbes are the main reasons for reducing aquaculture productivity. Hence, there is an immense need for the discovery of alternative and efficient treatment for quick recovery of diseased fishes. In the present study, Suaeda maritima leaf extracts (hexane, diethyl ether, ethanol, and water) were screened for in vitro and in vivo antibacterial and antioxidant activities. Out of all the four extracts, ethanolic extract showed highest antibacterial activity against S. aureus (4.9 ± 1.3 mm), B. subtilis (1.6 ± 0.3 mm), K. pneumoniae (4.2 ± 1.8 mm), and P. aeruginosa (4.1 ± 1.2 mm). Similarly, antioxidant activity was also higher for ethanolic extract (500 µg/mL) based on DPPH radical scavenging ability (71.6 ± 1.4%) and reducing potential (149 μg/mL) assays. Further, ethanolic extract was purified consecutively via column chromatography and preparative TLC where an active fraction was selected based on highest antibacterial (10.1 ± 1.4 mm) and antioxidant properties (82.3 ± 2.8%). Active fraction was loaded onto mass spectroscopy and identified the presence of four active constituents such as 1,2,9,10-tetramethoxy-6-methyl-5,6,6a,7-tetrahydro-4H-dibenzo[de,g]quinolin-3-yl) methanol; 3',7-Dimethoxy-3-hydroxyflavone; Saponin and (19R)9acetyl19hydroxy10,14dimethyl20oxopentacyclo[11.8.0.0 < 2,10 >.0 < 4,9 >.0 < 14,19 >]henicos-17-yl-acetate. Besides, in vivo studies were conducted on Catla catla fingerlings infected with P. aeruginosa under laboratory conditions. The fingerlings were segregated into 5 groups, among which group 4 and 5 were treated with crude and purified extracts. Both the extracts were efficient in treating infected fingerlings and recorded 100% survival rate which is even better than group-3 treated with a synthetic antibiotic (77%). Hence, S. maritima leaf extract can be considered as a possible alternative medicine in aquaculture.