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Growth Response of Spirulina Platensis in Papaya Skin Extract and Antimicrobial Activities of Spirulina Extracts in Different Culture Media

Nasima Akhtara
Bangladesh Council of Scientific and Industrial Research (BCSIR), Dhaka, Bangladesh.

Monzur Morshed Ahmed
Bangladesh Council of Scientific and Industrial Research (BCSIR), Dhaka, Bangladesh.

Nishat Sarker
Department of Microbiology, Stamford University, Dhaka, Bangladesh.

handaker Rayhan Mahbub
aBangladesh Council of Scientific and Industrial Research (BCSIR), Dhaka, Bangladesh and bDepartment of Microbiology, Stamford University, Dhaka, Bangladesh.

Md. Abdul Matin Sarker
Bangladesh Council of Scientific and Industrial Research (BCSIR), Dhaka, Bangladesh.

Abstract/ Executive Summary:

Growth response of Spirulina platensis in papaya skin extract media and their antimicrobial activity were studied. Five different concentrations e.g. 10gm/L, 8gm/L, 6 gm/L, 4 gm/L and 2gm/L of Papaya (Carica papaya) skin extract media and BD1 (control) medium were used in this study. After 8 days of cultivation, the optical density (0.33) was recorded in BD1 medium and among the five different concentrations of papaya skin extract media the maximum was found (0.31) in 6gm/L. Antimicrobial activity of Spirulina platensis grown in three media namely Zarrouk , BD1 media and media made from papaya skin extract was also studied. Only freeze dried Spirulina platensis powder extract showed inhibitory effect against bacteria and no antifungal activity was observed.

Keywords: Growth response, Antimicrobial activity, Papaya skin extract media, Spirulina platensis

Location:

Start Date:

End Date:

Program Area: Postharvest and Agro-processing

Commodity: Papaya

Objectives:

To develop a cheaper medium by utilizing locally available vegetable wastes such as skin of green papaya (Carica papaya) which contains necessary nutrients and mineral elements (Oloyede, 2005) which may be useful for growth of Spirulina platensis.

Methodology:

Media preparation: Fresh green papaya (Carica papaya) skin was peeled. Papaya skin was weighed and washed with tap water. 29 gm papaya skin was blended in 200 ml tap water. After blending, extract was sieved with 200μ mess cloth and stored at +4oC until use. For preparation of different concentration of papaya skin extract media, tap water was added separately with stored extract to make 10gm/L, 8gm/L, 6 gm/L, 4 gm/L and 2gm/L concentration and thoroughly mixed with 200 mg/L urea, 5gm/L NaCl and 3gm/L NaHCO₃ to increase the concentration of nitrogen, salinity and pH respectively. BD1 medium (Jahan et al., 1994) was used as control for this experiment. Each experiment was done with 3 replicatesInoculum preparation and maintenance Culture was carried out in 1L conical flask. 500 ml of papaya skin extract medium was given in each flask. Stock culture (Spirulina platensis) was collected from Biological Research Division, BCSIR, Dhaka which was maintained in Zarrouk (1966) medium. Equal amount of inoculums (20ml/L) was added into each flask (initial OD 0.11). The flasks were shaken everyday in the morning and evening. These were kept in a room near the window, exposed to natural condition. The optical density (OD) of the culture was recorded by Spectrophotometer (Type-Helios Gamma, NC-9423 UVG 1702E) for the maintenance of the growth. PH, temperature, light intensity, salinity and dissolved oxygen level of the culture were recorded. The condition of Spirulina culture was observed under compound microscope once a week and recorded. Sample preparation for screening of antimicrobial activity S. platensis was cultivated under photoautotrophic growth conditions. Zarrouk (1966), BD1 (Jahan, et al., 1994) and newly made Papaya Skin Extract media were used for cultivation. The initial pH was adjusted to 9.5. The culture temperature was maintained at 30° ± 0.1 °C. S. platensis was collected after 8 days of culture and dried in 3 different ways - freeze dry, sun dry and shade dry. Preparation of various extracts of S. platensis Freeze-dried, sun dried and shade dried S. platensis samples were mixed well with different solvents separately at the ratio of 0.5:10 w/v. Three different solvents (ethanol, methanol and chloroform) were used for the preparation of extracts of S. platensis. Twenty gram dried Spirulina powder was steeped separately in methanol and chloroform, 20gm dried powder was steeped in ethanol, then all the samples were kept for 72 hours at room temperature. Soaked mixtures were filtered (using What man filter paper-I).Then the filtrate was subjected to rotary vacuum evaporator (STUART, RE3022C) at 50º C and concentrated to gummy materials under reduced pressure. The gummy materials were then collected in small vials and then dried in room temperature. Thus crude extracts were obtained. The extracts were kept at +4 °C until use. The gummy extracts were dissolved in dimethyl sulfo-oxide (DMSO) prior to use in antimicrobial activity test.Determination of antibacterial and antifungal activities In vitro antibacterial studies were carried out against four bacterial pathogens viz Salmonella typhi CRL.(ICDDR.B), Escherichia coli ATCC 25922, Staphylococcus aureus ATCC 25923 Bacillus cereus BTCC 19 and two fungal pathogens viz Aspergillus fumigatus DSM 819 and Candida albicans ATCC 10239 which were obtained from the Microbiology Laboratory of IFST, BCSIR, Dhaka. Bacterial inoculums were prepared by Clinical and Laboratory Standards Institute (CLSI) guideline. Bacterial cultures were emulsified in normal saline and turbidity was matched with 0.5 McFarland turbidity standard. The agar cup method (Barry, 1980) was followed to investigate the antibacterial activity of the extracts. Wells of 6 mm diameter was punched over the agar plates using a sterile cork borer. The bottoms of the wells were sealed by pouring 50 - 100 μl of molten MHA into the scooped out wells. Using a micropipette, extracts of different solvents were added to different wells in the inoculated plate. These plates were then kept at 4ºC for 2-4 hours and then incubated at 37 ºC for 24 hours. Inhibition zones around the wells confirm the antibacterial activity of the respective extracts. The poisoned food technique (Grover and Moore, 1962) was used to screen antifungal activity. 0.1 ml extract of Spirulina platensis in respective solvent was taken by sterilized pipette in a sterile petriplate and then 20 ml of Sabouraud dextrose agar medium was poured into the petriplate, mixed well and allowed to solidify. Inoculation was done at the centre of each plate with 5 mm mycelium block for fungus. The mycelium block was prepared with the help of cork borer from the growing area of a 5 days old culture of the test fungi on sabouraud dextrose agar. The inoculated plates were incubated at 25ºC ±2 for 3 to 5 days. After 5 days of incubation the diameter of fungal radial mycelia growth was measured. The average of three measurements was taken as redial mycelial growth diameter of the fungus in mm. The percentage inhibition of mycelia growth of the test fungus was calculated by following method:

Source of Information: Bangladesh J. Sci. Ind. Res. 47(2), 147-152, 2012

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Achievement/Findings:

The present study suggests that vegetable wastes can be used as a good source of nutritive media for Spirulina culture at domestic level. Active ingredients present in S. platensis have diverse biological activity. Identification of these active ingredients present in S. platensis may be interesting topic to study antimicrobial activity of S. platensis

Knowledge Management: Journal