Background: Multidrug resistance of microbes forced the scientific community to search for newer antibiotics to treat infectious diseases. The literature review revealed that fishermen used the sea cucumber species for healing the wounds. Aim: As sea cucumber is rich in microbial flora, an attempt has been made to screen the antibiotic-producing microbes from them. Materials and Methods: Five sea cucumbers were collected from Kanyakumari district, Tamil Nadu, and dissected. The intestinal fluids and coelomic fluids were collected and named as IF and cystic fibrosis CF, respectively. The crowded plate method was followed to screen the antibiotic-producing microbes. As part of the primary screening, the perpendicular streak method was carried out to reveal the broad-spectrum potential of isolates. The selected isolates which showed the broad spectrum of activity were grown in soyabean casein broth media to produce the antibiotic principles. After the separation of cells from the fermented media, the agar well method was carried out against test organisms to reveal the broad spectrum of activity of the fermented broths. Results: The present investigation resulted in the isolation of 35 isolates with antibiotic-producing ability in crowded plate method. Out of 35 isolates, only 9 of them passed the primary screening. The secondary screening revealed that the fermented broths of three isolates (named IF 32 , IF 52 , and CF 42) were found to have a better broad spectrum of activity.
Introduction: Three broad-spectrum antibiotic-producing microbes (named IF 32 , IF 52 , and CF 42) were identified and isolated from intestinal region and coelomic fluids of sea cucumber species collected from Kanyakumari district, Tamil Nadu. Biochemical characterization of the isolated microbes and optimization of culture characteristic (for maximum antibiotic productivity) were performed. The present study focuses on extraction and purification of antibiotic principles from the culture medium of the selected isolates. Methodology: The antibiotic fermentation was carried out in the optimized fermentation medium under optimum conditions separately for the isolated species. After separation of the cells by filtration, the fermented culture media were subjected to solvent extraction using 1-butanol, chloroform, ethyl acetate, and hexane. Obtained residues were subjected to antibiotic screening against Staphylococcus aureus (MTCC 1430) by disc plate method. Purification of the selected residues which showed antibiotic activity was done by column chromatography using silica gel as packing material. Collected fractions of the three antibiotic residues were subjected to bioautography to ensure the purification of the antibiotics. Results: The results show that chloroform, 1-butanol, and ethyl acetate are found to be suitable solvents for the extraction of antibiotics from the fermented cultures of IF 32 , IF 52 , and CF 42 , respectively. Bioautography performed using fractions obtained from column chromatographic separation ensured the purification of the antibiotic principles.
Introduction: Five sea cucumber species were collected from fishermen at coastal region of Kanyakumari district, Tamil Nadu, and dissected aseptically. Intestinal fluids and coelomic fluids were collected. After serial dilution of the collected fluids, primary screening was carried out by crowded plate method using modified Soybean Casein Agar media (Kester et al.). Selected antibiotic-producing organisms producing zone of inhibition (ZOI) higher than 6 mm (ZOI >6 mm) were evaluated for broad spectrum of activity using two Gram positive, two Gram negative, and two fungal species by perpendicular streak method. Biochemical characterization and nutrient optimization were carried out for selected three isolates (named IF 32 , IF 52 , and CF 42) which were found to have broad spectrum of activity as well as passed Kirby-Bauer antimicrobial susceptibility test. This article focuses on optimization of temperature, pH, and duration for maximum antibiotic productivity of the selected isolates. Methodology: Optimum temperature, pH, and duration of maximum antibiotic production were evaluated for the selected antibiotic producers. Crude antibiotic was collected after 24 h of incubation under different temperature and pH conditions and tested against Staphylococcus aureus (MTCC 1430). The specific temperature and pH at which maximum ZOI was produced by the collected crude extracts against test organisms were considered. The duration of maximum antibiotic production was evaluated by incubating the isolated organisms under the optimized temperature and pH. Results: The optimum temperature for the production of crude antibiotic from IF 32 , IF 52 , and CF 42 was found to be 32°C, 35°C, and 31°C, respectively. The pH at which maximum antibiotic production was observed is found to be 8, 7, and 9, respectively, for IF 32 , IF 52 , and CF 42. The duration of maximum antibiotic productivity at the optimized conditions was found to be 4 th , 5 th , and 3 rd days, respectively, for IF 32 , IF 52 , and CF 42 .
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