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Callyspongia Crassa and C. Siphonella (Porifera, Callyspongiidae) As a Potential Source for Medical Bioactive Substances, Aqaba Gulf, Red Sea, Egypt Hassan A

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Callyspongia Crassa and C. Siphonella (Porifera, Callyspongiidae) As a Potential Source for Medical Bioactive Substances, Aqaba Gulf, Red Sea, Egypt Hassan A Ibrahim et al. The Journal of Basic and Applied Zoology (2017) 78:7 The Journal of Basic DOI 10.1186/s41936-017-0011-5 and Applied Zoology RESEARCH Open Access Callyspongia crassa and C. siphonella (Porifera, Callyspongiidae) as a potential source for medical bioactive substances, Aqaba Gulf, Red Sea, Egypt Hassan A. H. Ibrahim1, Hussein A. El-Naggar2*, Khalied A. El-Damhougy2, Mansour A. E. Bashar2 and Fekry M. Abou Senna2 Abstract Background: Sponges are champion of bioactive producers because of the variety of products that have been found from them. Most bioactive compounds extracted from sponges were classified into antibiotic, antiviral, antitumor, and anti-inflammatory. Tow marine sponge species were collected during winter 2016 from Gulf of Aqaba, Red Sea, Egypt viz., Callyspongia crassa and Callyspongia siphonella. The collected sponge species belong to family Callyspongiidae. The sponge samples were extracted by ethanol and investigated as a promising source of natural products which can be used as antitumor, antiviral, and antimicrobial, antioxidant, and anti-inflammatory agents. Results: Results revealed that the crude extract of C. siphonella showed high antitumor activity with value of 5.57 and 1.39 μg/ml as IC50 against colon cancer (caco-2) and breast cancer (Mcf-7) on cell line, respectively. Also the C. crassa extract showed high cytotoxic effect to Vero cell with HAV (hepatitis A virus), which exhibits that MIC was 9.765 μg/ml, the most effective extract was of C. crassa against Escherichia coli, Pseudomonas aeruginosa,andCandida albicans. Also, ethanolic crude of C. siphonella showed positive antibacterial activity against P. aueroginosa. Indeed, there much other extracts exhibited no antibacterial activities, especially all extracts against Staphylococcus aureus and Vibrio damsela. The C. crassa extract conducted an antioxidant activity in corresponding to ascorbic acid standard with value of 671 μg/ml as IC50. There was only one crude extract that had anti-inflammatory activity; it was of C. crassa (61.47%). Conclusions: The present investigation confirmed that the crude extract of C. siphonella conducted to promising antitumor agent against colon and breast cancer, while the crude extract of C. crassa conducted to promising anti-inflammatory and antiviral agents. In summary, the marine sponges are an indispensable, chemodiverse, biodiverse, and rich source of natural products and secondary metabolites with potent pharmacological effects. The studies should be interested with sponge- derived bioactive compounds as a promising strategy that deserves further attention in future investigations in order to address the limitations regarding sustainable supply of marine drugs. Keywords: Sponges, Red Sea, Antitumor, Antiviral, Antimicrobial, Anti-inflammatory * Correspondence: [email protected] 2Faculty of Science, Al-Azhar University, Cairo, Egypt Full list of author information is available at the end of the article © The Author(s). 2017 Open Access This article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. Ibrahim et al. The Journal of Basic and Applied Zoology (2017) 78:7 Page 2 of 10 Background Therefore, the present study aims to extract bioactive sub- Marine organisms are rich source of structurally novel and stances from marine sponges species inhabiting Gulf of biologically active metabolites. Some studies have suggested Aqaba, Red Sea, Egypt in a hope to be used as potential that some bioactive compounds isolated from marine or- medical agents in several applications (such as; antiviral, anti- ganisms have been shown to have anticancer, anthelmintic, microbial, antioxidant, antitumor, and anti-inflammatory). antibacterial, antifungal, antiviral, antimalarial, anticoagu- lant, antiprotozoal, antituberculosis, or anti-inflammatory Methods and other pharmacological activities (Mayer and Hamann Sampling and identification of sponge specimens 2005; Blunt et al. 2005; Somnath and Ghosh 2010). Many The investigated specimens were collected during winter bioactive compounds have been obtained from various 2016 at different depths along Gulf of Aqaba by SCUBA marine animals like sea hares, nudibranchs, bryozoans, diving. The identification of all specimens has been care- tunicates, sponges, soft corals, sea slugs and marine micro- fully checked on the basis of morphological characters organisms, etc (Donia and Hamann 2003). Among marine according to Systema Porifera (Hooper and Van Soest invertebrates, Porifera (sponges) remain the most prolific 2002) and the recent update undertaken in the World phylum, concerning novel pharmacologically active com- Porifera Database (Van Soest et al. 2008). pounds (Faulkner 2000). Richter (1907) outlined that the active component of the roasted bath sponge, used by Roger against Struma, is iodine. The work on sponge nat- Preparation of different sponge crude extracts ural products was systematically started by who isolated One hundred grams of each marine sponge specimen three nucleosides from the Caribbean sponge Cryptotethya- were macerated with 200 ml of 70% aqueous ethanol. crypta laubenfels. Antiviral properties of these nucleosides After soaking for a week with genital shaking, they were were demonstrated later (Privat et al. 1964) and initiated filtered through Whatman 542 filter paper. Solvent was the synthesis of analogues which led to the first antiviral evaporated using rotary evaporator to obtain soluble compound Ara-A (active againstherpesvirus)andantitu- extracts (Ballantine et al. 1987). mor compound Ara-C (effective in acute lymphoid leukemia). The compounds Ara-A and Ara-C are the only Antimicrobial assay marine invertebrate-related compounds in clinical use Microbial indicator strains (Guyot 2000). Arabinosyl Cytosine (Ara-C) is currently sold The bacterial indicators were Escherichia coli, Pseudomonas by the Pharmacia & Upjohn Company under the brand aeruginosa, Staphylococcus aureus, Vibrio damsela,and name Cytosar-UR. Since this pioneering work, number of Candida albicans. novel bioactive molecules have been discovered from this group, which include cytotoxins, antibiotics, antiviral, and Media and bacterial cultures anti-inflammatory compounds. It is known that sponges −1 the most diverse marine invertebrates have evolved antag- Nutrient broth (Atlas 1997): composed of (gl )the onistic effects against other invading organisms, which following: yeast extract, 2; beef extract, 1; peptone, 5; – involves the production of the secondary metabolites (Wah sodium chloride, 5. Agar (15 20) was added for obtaining et al. 2006). The secondary metabolites isolated from mar- nutrient agar. All bacterial pathogenic strains were main- inespongesmaybealternativesourceforvectorcontrol tained on nutrient agar slants. Bacterial inoculate were agents to replace existing and highly toxic synthetic insecti- prepared by inoculating 100 ml of nutrient broth medium, cides and will play an important role in future insecticide and incubated shaken (250 rpm) at 30 °C for 24 h until development program (Rao et al. 2008). Initial experimental late logarithmic phase of growth (A550 = 1). Antibacterial results suggested that sponges have the potential to provide activity was carried out on nutrient agar plates. future drugs against important diseases, such as cancer, a range of viral diseases, malaria, and inflammations (Perdi- Antimicrobial activity of sponge crude extracts caris et al. 2013). Also, some of the most potential sponge- The well-cut diffusion technique was used to test the ability derived bioactive molecules include the anti-inflammatory of the crude extracts of sponges to inhibit the growth of compound Manolide from the Palauan sponge Luffariella indicator bacteria and Candida. One hundred microliters variabilis (De-Silva and Scheuer 1980), the immunosup- of the tested crude extracts were transferred into each inoc- pressive and cytotoxic compound Discodermolide from the ulated well after sterilizing by ultra-filtration. All plates were deep sea sponge Discodermia dissolute (Gunasekera et al. incubated at appropriate temperature for 24–48 h. After 1990), anti-angiogenic brominated compound aeroplysina- incubation period, the radius of clear zone around each well 1fromaspongeAplysina aerophoba (Rodriguez-Nieto (Y) and the radius of the well (X) were linearly measured in et al. 2002), tubulin polymerizing compound dictyostatin-1, mm, where dividing Y2 over X2 determines an absolute unit a polyketide from Spongia sp. (Isbrucker et al. 2003) etc. (AU) for the clear zone. The absolute unit of crude extract, Ibrahim et al. The Journal of Basic and Applied Zoology (2017) 78:7 Page 3 of 10 which indicates a positive result, was calculated according The percentage cell viability was calculated using the to the equation (Yang et al. 1992): Microsoft Excel ®. Percentage cell viability was calculated as follows: 2 2 AU ¼ Y π=X π Cell viability% ¼ Mean Abs control–Mean Abs plant extract Â100 Mean Abs control, where Abs absorbance at 490 nm Antitumor assay Cultures were viewed using an inverted microscope to Antiviral assay assess the degree of confluence and the absence of bacter- Vero cells used in the present study were established by ial and
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