Antifungal Activity of Ninamental Gland and Acessory Ninamental Gland Extracts of Squid and Cuttlefish

Abstract- Nidamental gland complex formed a part of visceral mass in cephalopods viz., squid and cuttlefish. Accessory nidamental glands are located at the anterior end of the nidamental glands, closely associated with ventral surface of ink sac. Visceral mass from mature ripe females of squid (Uroteuthis singhalensis) and cuttlefish (Sepia pharaonis) were collected during the month of October, 2012 to December, 2012 which is the peak season of maturity. The nidamental glands (NGs) and accessory nidamental glands (ANGs) of squid and the cuttlefish were dissected free and removed aseptically. The ANGs were conspicuously coloured only in sexually mature females.

The ANGs and NGs were extracted in ethanol and butanol by the exhaustive method (homogenized ripe gland tissue 2.5 g with 5.0 ml butanol and ethanol) and stored at -20 ̊ C for further antifungal assay. The ANGs of mature female squid and cuttlefish appear red or mottled red and orange in colour and the extract is reddish colour and very pale orange in colour. While NGs of both the species appears as creamy white in colour and the extract is colourless. The occurrence of these colours in ANGs has been attributed to pigments of symbiotic bacteria.

Antifungal activity of the ethanol extracts of ANG and NG from the mature female of cuttlefish (Sepia pharaonis) and squid (Uroteuthis singhalensis) against the four fungal strains of Candida albicans, Aspergillus flavus, Aspergillus fumigates and Fusarium solani were studied. The four fungal species were procured from the Institute of Microbial Technology, Chandigarh. The ANG and NG ethanol extracts of cuttlefish and squid showed good antifungal activity against the fungus species of Aspergillus fumigates with 15mm, 12mm and 12mm respectively of inhibition halos. The ethanol extracts of cuttlefish and squid ANG and NG showed moderate antifungal activity against the fungus species of Candida albicans with the formation of growth inhibition halos of 10mm each. The ANG and NG ethanol extracts of squid and cuttlefish did not show any antifungal property against the fungus species Aspergillus flavus and Fusarium solani. The negative did not show any antifungal activity against the four fungal species.

The antifungal activity of the ANG and NG butanol extracts of the cuttlefish and squid species were studied against four fungal micro-organisms viz. Candida albicans, Aspergillus flavus, Aspergillus fumigates and Fusarium solani. The ANG and NG butanol extracts of the cuttlefish and squid showed potential antifungal property against the fungal species of Aspergillus flavus (17 mm and 16 mm and 15 mm and 15mm respectively of growth inhibiting halos). The antifungal activity of the ANG and NG butanol extracts of the squid against the fungal species of Aspergillus fumiagatus showed good antifungal activity with 14 mm and 13 mm as growth inhibition halos. While the butanol extracts that of cuttlefish did not show any antifungal activity. The ANG and NG butanol extracts of cuttlefish and squid showed moderate to good antifungal activity (10 mm, 10 mm, 10 mm and 15 mm respectively diameter of the growth inhibition halos). The ANG and NG butanol extracts of cuttlefish and squid did not show any antifungal activity against the fungal species of Fusarium solani. The negative control did not show any antifungal property against the four fungal species.

Based on previous results on the biochemical composition of ANGs of S. pharaonis and U. singhalensis from the Cochin area, it is concluded that the major fatty acids like palmitic acid, DHA, Oleic acid and EPA present in the glands may be the bioactive substances responsible for the antifungal activity.

ANG and NG of squid and cuttlefish play an important role in protecting eggs and embryos due to their antimicrobial (antibacterial and antifungal) properties. Much information is available on the antibacterial properties of the gland extracts but this is the first study, which brings out the antifungal activity of ANG and NG extracts of squid and cuttlefish.