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Helicobacter pylori were considered one of the most bacterial infections in the world. Recently, the resistance of antibiotic against H. pylori is increasing and hence, it is necessary to find new and natural effective agents. Marine organisms and microorganism produce complex secondary metabolites and some of these compounds are antimicrobials agents. This study aimed to provide preliminary data to determine if marine Hyrtios erectus sponge, actinomycetes, and fungi extracts can inhibit the growth of H. pylori. In this study, the methanol extracts of the three samples of Hyrtios erectus sponge and the ethyl acetate extracts of seven actinomycetes strains and four fungi strains were used to determine the antibacterial effect against a standard strain of H. pylori. The samples of actinomycetes and fungi were identified by using 16S rRNA amplification for actinomycetes and Internal Transcribed Spacer-PCR for fungi using Genetic Analyzer 3500, and then the DNA sequences were compared through BLAST and alignment with other actinomycetes and fungi which already present in the GenBank from various study areas. Results indicated that the anti-H. pylori screening showed that only 13 from 14 extracts exhibited anti-H. pylori effect. Among them, the methanol extract of Hyrtios erectus (1), the ethyl acetate extract of actinomycete strain S7 and the ethyl acetate extracts of fungi strains M1, M2, M3, M4 exhibited strong effect against H. pylori more than other extract with minimal inhibitory concentrations 3.9, 15.63, 3.9, 15.63, 3.9 and 7.81 µg/ml, respectively. The 16S rRNA and ITS rRNA gene sequences and phylogenetic analysis provided strong evidence for the isolates of actinomycetes and fungi, respectively. The actinomycetes were identified as seven species that belong to Streptomyces genus. The fungi isolates were identified as three species that belong to Aspergillus genus and one species that belong to Penicillium genus.
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