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Journal of Drug Delivery and Therapeutics

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Antibacterial Activity of Haematococcus pluvialis Crude Astaxanthin Extract

*Ashaq Hussain Rather, Surendra Singh, Sameer Choudhary

 Algal Biotechnology Laboratory, Department of Post Graduate Studies and Research in Biological Sciences, Rani Durgavati University, Jabalpur-482001, (M.P.), India

1. INTRODUCTION

The growing demand in the cosmetic formulation with natural ingredients continues to raise and now-a-days this has been a trend to replace the synthetic preservatives with natural ones. The natural preservatives are preferred from plants, bacteria, fungi, and algal sources. Astaxanthin is a xanthophyll carotenoid¹. The main biological sources of astaxanthin include crustacean crustacean extracts, Haematococcus pluvialis, yeast Rhodotorularubra, and Phaffia rhodozyma. The aforementioned natural sources cannot compete with the synthetic products available ². Haematococcus pluvialis, freshwater green microalga is considered a potent producer of pigment astaxanthin ³. In our previous study ^{4, 5} H. pluvialis was cultivated and astaxanthin was extracted from the green alga H. pluvialis. In present study, efforts have been made to study antibacterial activity of Haematococcus pluvialis crude astaxanthin extract.

2. MATERIALS AND  METHODS
   • Cultivation

Haematococcus pluvialis culture was obtained from culture collection of algae at the University of Texas, Austin, USA. H. pluvialis culture was grown in the bold basal medium (BBM) ⁶. The astaxanthin accumulated cells, were harvested, dried in the oven at 70 ^oC ⁷.

• Preparation of Samples

10 mg biomass of H. pluvialis was mixed in mortar and pestleand extracted with acetone, methanol, DMSO and hexane, centrifuged at 2800x g for 10 min at 4 ^oC, the pellet was discarded and the supernatant was taken for the estimation of astaxanthin. The absorbances of the extracts were determined at 492 nm and the amount of the pigment was quantitatively measured according to Davies ⁸. Acetone was evaporated in the rotary evaporator. The crude astaxanthin extracts were individually tested for Escherichia coli, Salmonella typhi, Vibrio cholera and Staphylococcus aureus. The antibacterial activity was tested using disc diffusion method. 1 ml of test organism at concentration of 10^-5 cells mL^-1 was spread on Mueller-Hinton agar plate. 10 μl astaxanthin, extracted with acetone, methanol, DMSO and hexane was poured in the wells. The plates of bacterial strain were incubated at 37 °C for 24. The inhibition zone discs were observed and the diameters of clear zones were measured. The bacteria strains were sub cultured and maintained on nutrient agar (Beef extract 3.0 g, Peptone 5.0 g, NaCl 5.0 g, Agar 15.0 g, pH 7.4, dissolved in one liter distilled water and sterilized at 121 ^oC for 15 minutes).

3. RESULTS AND DISCUSSION
   • Cultivation

Haematococcus pluvialis was grown in the bold basal medium for 30 days and the cells were harvested and astaxanthin was extracted with different solvents (Figure 1)

Figure 1: Haematococcus pluvialis (A) green stage and red stage (B) astaxanthin extract

Astaxanthin has been extracted with acetone, methanol, DMSO and hexane and the crude astaxanthin was tested for the E.coli, Salmonella typhi, Vibrio cholera, Staphylococcus aureus. The results showed that the green algae H. pluvialis crude extract (10μl) possess antibacterial activity by showing the inhibition zone around the discs. Highest antibacterial activity was found as 10.2 ±0.20 mm extracted with acetone on Escherichia coli. In methanol, DMSO and hexane   extracts, highest antibacterial activity was found as 8.8±0.12, 9.4 ±0.15 and 8.7 ±0.12 mm on Salmonella typhi, Salmonella typhi, Staphylococcus aureus respectively as seen in Table 1.

Table 1: Antibacterial activity of crude H. pluvialis astaxanthin

The antioxidant activities of astaxanthin are better than β-carotene and vitamin-E. This is the reason due to which makes it protects against inflammation, UV radiation, aging and skin cancer. Astaxanthin is used in cosmetic, food and feed industries. H. pluvialis astaxanthin is found effective agent for the regulation and maintenance of uric and hepatic enzymes in rat model animals ⁹. The pharmacokinetic data has revealed that a single dose of 10 mg astaxanthin can remain in the human blood for a day and 100 mg for approximately three days ¹⁰. A dose of 1 mg when given once daily for a month can increase the blood levels ¹¹. The animal study showed that astaxanthin higher than 120 mg a day in human ¹², revealed no harmful effects. Gonzalez et. al ¹³ observed that extract of green algae have umpteen antimicrobial activities against B. subtilis and S. aureus. The green algae extract showed potential effects against Gram-positive bacteria ¹⁴.

4. CONCLUSION

The current results revealed that crude astaxanthin extracts prevented the bacterial activity hence H. pluvialis astaxanthin may be consider as a preservative in different food formulations.

ACKNOWLEDGEMENT

We are thankful to the head, department of post graduate studies and research in biological sciences, Rani Durgavati University, Jabalpur-482001, (M.P.) India for providing necessary facilities.

CONFLICT OF INTEREST

We declare that we have no conflict of interest.

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