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Biotoxicological Assay of Selected Philippine Poisonous Crab
Author(s) -
Atanacio Francis Rainer Barruga
Publication year - 2016
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.30.1_supplement.1192.4
Subject(s) - portunus pelagicus , bacillus cereus , antimicrobial , brine shrimp , biology , staphylococcus aureus , microbiology and biotechnology , staphylococcus epidermidis , bacteria , zoology , fishery , crustacean , genetics
Ten selected poisonous crabs namely: Dromidiopsis dormia, Calappa calappa, Charybdis notator, Carpilius maculatus (Adult), Carpilius convexus, Atergatis intgerrimus, Lophozozymus pictor, Demania sp., Banareia sp. and Portunus pelagicus were used in antimicrobial assay and brine shrimp lethality assay. The most bioactive extract of Lophozozymus pictor was fractionated using gel filtration chromatography. The fractionated samples were freeze dried, lyophilized, and were subjected again to biotoxicological assay with the same test organisms. The results showed that the most biotoxic fractions of Lophozozymus pictor were F11–13 which exhibited antimicrobial inhibitory zones of 23 mm, 23 mm, 21 mm and 28 mm against Staphylococcus aureus, Bacillus cereus, Staphylococcus epidermidis, and Epidermophyton floccosum respectively. These indicate that Lophozozymus pictor contains antimicrobial principles which is bioactive against pathogenic microorganisms. The results in the brine shrimp lethality assay affirmed that selected Philippine crabs contain biotoxic compounds. I Bacterial Inhibitory Zones of (in millimeter) of Dromidiopsis dormia, Calappa calappa, Charybdis notator, Carpilius maculates (Adult), Carpilius convexus, Lophozozymus pictor, Atergatis integerrimus, Demania sp. Carpilius maculatus (Juv,),, Banareia sp., and Portunus pelagicus (Edible) extracts, Tetracycline, and Distilled Water against Gram‐Positive Bacteria after 24 hours of incubation.Gram‐Positive Bacteria Staphylococcus aureus (004) Bacillus cereus (011) Staphylococcus epidermidis(024)I II III m I I III m I II III MCarpilius maculates (Adult) 0 0 12 4 0 0 0 0 0 9 9 6Lophozozymus pictor 10 7 0 5.7 10 8 10 9.3 9 11 7 9Dromidiopsis dormia 0 0 13 5.7 12 12 16 13.3 9 9 10 9.33Demania sp. 0 0 0 0 0 12 9 7 0 10 11 7Charybdis notator 0 0 0 0 0 0 0 0 0 0 0 0Calappa calappa 0 0 0 0 0 0 0 0 0 0 0 0Atergatis integerrimus 0 0 0 0 0 0 0 0 0 0 0 0Carpilius conuexus 0 0 0 0 0 0 0 0 0 0 0 0Carpilius maculatus (Juv.) 0 0 0 0 0 0 0 0 0 0 0 0Banareia sp. 0 0 0 0 0 0 0 0 0 0 0 0Portunus pelagicus (Edible) 0 0 0 0 0 0 0 0 0 0 0 0Tetracycline 14 14 14 14 8 8 8 8 11 14 10 11.7Distilled Water 0 0 0 0 0 0 0 0 0 0 0 0II Bacterial Inhibitory Zones of (in millimeter) of Dromidiopsis dormia, Calappa calappa, Charybdis notator, Carpilius maculates (Adult), Carpilius convexus, Lophozozymus pictor, Atergatis integerrimus, Demania sp. Carpilius maculatus (Juv.)„ Banareia sp., and Portunus pelagicus (Edible) extracts, Tetracycline, and Distilled Water against Gram‐Positive Bacteria after 48 hours of incubation.Gram‐Positive Bacteria Staphylococcus aureus (004) Bacillus cereus (011) Staphylococcus epidermidis (024)I II III M I I III m I II III mCarpilius maculates (Adult) 0 0 13 4.3 0 0 0 0 0 9 10 6.3Lophozozymus pictor 10 9 11 10 10 0 0 3.3 10 11 0 7Dromidiopsis dormia 0 0 14 4.7 10 11 0 7 9 10 10 9.7Demania sp. 0 0 0 0 0 10 0 3.3 0 12 11 7.7Charybdis notator 0 0 0 0 0 0 0 0 0 0 0 0Calappa calappa 0 0 0 0 0 0 0 0 0 0 0 0Atergatis integerrimus 0 0 0 0 0 0 0 0 0 0 0 0Carpilius convexus 0 0 0 0 0 0 0 0 0 0 0 0Carpilius maculatus (Juv.) 0 0 0 0 0 0 0 0 0 0 0 0Banareia sp. 0 0 0 0 0 0 0 0 0 0 0 0Portunus pelagicus (Edible) 0 0 0 0 0 0 0 0 0 0 0 0Tetracycline 14 14 14 14 8 8 8 8 11 14 10 11.7Distilled Water 0 0 0 0 0 0 0 0 0 0 0 0III Bacterial Inhibitory Zones of (in millimeter) of Dromidiopsis dormia, Calappa calappa, Charybdis notator, Carpilius maculates (Adult), Carpilius convexus, Lophozozymus pictor, Atergatis integerrimus, Demania sp. Carpilius maculatus (Juv.),, Banareia sp., and Portunus pelagicus (Edible) extracts, Tetracycline, and Distilled Water against Gram‐Negative Bacteria after 24 hours of incubation.Gram‐Negative Bacteria Escherichia coli (002) Salmonella typhi (007) Klebsiella pneumoniae ()I II III m I I III m I II III mCarpilius maculates (Adult) 12 0 0 4 0 0 10 3.3 0 0 0Lophozozymus pictor 12 10 0 7.3 0 0 11 3.7 10 10 7Dromidiopsis dormia 12 10 0 7.3 0 12 9 7 8 0 0Demania sp. 12 0 0 4 0 0 0 0 7 0 0Charybdis notator 0 0 0 0 0 0 0 0 0 0 0Calappa calappa 0 0 0 0 0 0 0 0 0 0 0Atergatis integerrimus 0 0 0 0 0 0 0 0 0 0 0Carpilius convexus 0 0 0 0 0 0 0 0 0 0 0Carpilius maculatus (Juv.) 0 0 0 0 0 0 0 0 0 0 0Banareia sp. 0 0 0 0 0 0 0 0 0 0 0Portunus pelagicus (Edible) 0 0 0 0 0 0 0 0 0 0 0Tetracycline 14 14 14 14 8 8 8 8 11 14 10Distilled Water 0 0 0 0 0 0 0 0 0 0 0IV Bacterial Inhibitory Zones of (in millimeter) of Dromidiopsis dormia, Calappa calappa, Charybdis notator, Carpilius maculates (Adult), Carpilius convexus, Lophozozymus pictor, Atergatis integerrimus, Demania sp. Carpilius maculatus (Juv.),, Banareia sp., and Portunus pelagicus (Edible) extracts, Tetracycline, and Distilled Water against Gram‐Negative Bacteria after 48 hours of incubation.Gram‐Negative Bacteria Escherichia coli (2) (004) Salmonella typhi (007) Klebsiella pneumoniae (020)I II III m I I III m I II III mCarpilius maculates (Adult) 11 9 8 9.3 9 11 11 10.3 11 11 11 11Lophozozymus pictor 9 11 9 9.7 12 10 8 10 13 10 10 11Dromidiopsis dormia 9 10 9 .3 10 13 9 10.7 0 0 0 0Demania sp. 16 0 0 5.3 0 8 0 2.7 10 0 0 3.3Charybdis notator 0 0 0 0 0 0 0 0 0 0 0 0Calappa calappa 0 0 0 0 0 0 0 0 0 0 0 0Atergatis integerrimus 0 0 0 0 0 0 0 0 0 0 0 0Carpilius convexus 0 0 0 0 0 0 0 0 0 0 0 0Carpilius maculatus (Juv.) 0 0 0 0 0 0 0 0 0 0 0 0Banareia sp. 0 0 0 0 0 0 0 0 0 0 0 0Portunus pelagicus (Edible) 0 0 0 0 0 0 0 0 0 0 0 0Tetracycline 14 14 14 14 8 8 8 8 11 14 10 11.3Distilled Water 0 0 0 0 0 0 0 0 0 0 0 0V Antifungal Inhibitory Zones of (in millimeter) of Dromidiopsis dormia, Calappa calappa, Charybdis notator, Carpilius maculates (Adult), Carpilius convexus, Lophozozymus pictor, Atergatis integerrimus, Demania sp. Carpilius maculatus (Juv.),, Banareia sp., and portunus pelagicus (Edible) extracts, Nystatin, and Distilled Water against Fungi after 72 hours of incubation.Gram‐Negative Bacteria Candida albicans (2049) Epidermophyton floccosum (E) Fusarium moniliforme (F)I II III m I I III m I II III MCarpilius maculates (Adult) 0 0 13 4.3 16 10 20 15.3 22 10 26 19.3Lophozozymus pictor 0 0 0 0 20 16 18 18 34 22 26 27.3Dromidiopsis dormia 0 11 11 7.3 18 14 20 17.3 16 14 22 17.3Demania sp. 11 17 0 9.3 16 10 20 15.3 12 10 26 16Charybdis notator 0 0 0 0 0 0 0 0 0 0 0 0Calappa calappa 0 0 0 0 0 0 0 0 0 0 0 0Atergatis integerrimus 0 0 0 0 0 0 0 0 0 0 0 0Carpilius convexus 0 0 0 0 11 10 10 10.3 18 16 20 18Carpilius maculatus (Juv). 0 0 0 0 11 18 14 14.3 22 20 20 21.7Banareia sp. 0 0 0 0 8 9 8 8.3 10 0 0 3.3Portunus pelagicus (Edible) 0 0 0 0 0 0 0 0 0 0 0 0Nystatin 10 11 14 11.7 9 8 12 9.7 12 11 10 11Distilled Water 0 0 0 0 0 0 0 0 0 0 0 0VI Antifungal Inhibitory Zones of (in millimeter) of Dromidiopsis dormia, Calappa calappa, Charybdis notator. Carpilius maculates (Adult), Carpilius convexus, Lophozozymus pictor, Atergatis integerrimus. Demania sp. Carpilius maculatus (Juv.)„ Banareia sp.. and Portunus pelagicus (Edible) extracts, Nystatin, and Distilled Water against Fungi after 168 hours of incubation.Gram‐Negative Bacteria Candida albicans (2049) Epidermophyton floccosum (E) Fusarium moniliforme (F)I II III M I I III M I II III MCarpilius maculates (Adult) 0 0 0 0 10 11 16 15.3 12 16 13 13.7Lophozozymus pictor 0 0 0 0 20 11 8 18 18 12 12 14Dromidiopsis dormia 0 0 0 0 10 11 10 13.7 24 16 12 17.3Demania sp. 0 0 0 0 22 10 10 2.7 24 16 13 17.7Charybdis notator 0 0 0 0 0 0 0 0 0 0 0 0Calappa calappa 0 0 0 0 0 0 0 0 0 0 0 0Atergatis integerrimus 0 0 0 0 0 0 0 0 0 0 0 0Carpilius convexus 0 0 0 0 10 0 10 6.7 34 12 12 19,3Carpilius maculatus (Juv.) 0 0 0 0 10 14 10 11.3 19 15 15 16.3Banareia sp. 0 0 0 0 7 11 0 6 13 10 10 11Portunus pelagicus (Edible) 0 0 0 0 0 0 0 0 0 0 0 0Nystatin 10 11 14 11.7 9 8 12 9.7 12 11 10 11Distilled Water 0 0 0 0 0 0 0 0 0 0 0 0VII Mean biotoxicity of Dromidiopis dormia, Calappa calappa, Charybdis notator, Carpilius maculates (Adult), Carpilius convexus, Lophozozymus pictor, Atergatis integerrimus, Demania sp. Carpilius maculatus (Juv)., Banareia sp., and Portunus pelagicus (Edible) extracts at 20 μL, 18 μL, 16 μL, 14 μL, and 12 μL concentration by Brine Shrimp Lethality Assay using Artemia salina after 30 mins, 60 mins, and 90 mins.Concentration (μL) Carpilius maculates (Adult) Lophozozymus pictor Dromidiopis dormia Demania sp. Charybdis notator Calappa calappa Atergatis integerrimus Carpilius convexus Carpilius maculatus (Juv.) Banareia sp. Portunus pelagicus (Edible)After 30 mins. 12 15 15 15 15 15 15 15 15 15 15 1514 15 15 15 15 15 15 15 15 15 15 1516 15 15 15 15 15 15 15 15 15 15 1518 15 15 15 15 15 15 15 15 15 15 1520 15 15 15 15 15 15 15 15 15 15 15 After 60 mins. 12 9 14 12 8 8 14 7 8 8 14 1514 11 15 13 8 12 12 7 8 8 14 1316 11 15 12 12 12 14 3 14 8 14 1518 11 15 13 14 12 15 7 15 12 14 1520 11 14 14 13 15 14 3 12 14 8 15 After 90 mins. 12 9 8 12 8 8 8 5 8 8 8 1514 11 9 12 7 12 8 4 8 7 8 1316 10 12 12 12 12 12 2 13 7 12 1518 10 10 12 12 12 12 3 12 10 12 1520 9 10 14 13 8 12 3 11 12 10 13Atergatis integerrimus Trial I Trial I Trial II Mean1 11 13 9 11 2 7 6 12 8.33 3 9 7 10 8.67 4 8 8 11 9 5 9 7 9 8.33 6 9 9 8 8.67 7 9 9 7 8.33 8 6 5 9 6.67 9 5 6 6 5.67 10 2 4 6 4 11 3 6 7 5.33 12 4 2 7 4.33 13 3 8 4 5 14 2 2 5 3 15 4 5 3 4 16 6 6 5 5.67 17 8 7 9 8 18 5 4 5 4.67 19 9 5 6 6.67 20 10 8 6 8 21 8 9 8 8.33 22 6 8 9 7.67 23 9 9 4 7.33 24 12 8 6 8.67 25 9 9 8 8.67 26 7 7 8 7.33 27 10 7 8 8.33 28 12 6 6 8 29 9 6 5 6.07 30 11 12 7 10