Antagonistic properties of reef fish microflora

a technology of anti-antagonistic properties and reef fish, which is applied in the direction of antibodies to medical ingredients, paints with biocides, bacteria based processes, etc., can solve the problems of prolonging the life of drugs and inability to develop resistan

Inactive Publication Date: 2007-05-03
AEQUOR INC
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Benefits of technology

[0110] Reference organism (see Example 5) and fish isolate were cross streaked on top of the other on NA plate and incubated at 37° C. for 48 hours. Clearance zones of inhibited growth for reference organism were measured and recorded in mm as width of the inhibition streak band. A zone of clearance of 1.0 mm and larger indicates a positive result of inhibition.
[0111] 33.3% of the fish isolates tested showed antibacterial activity against two of the nine reference strains tested, S. aureus and S. epidermidis (Table 4). Five isolates (P5-1, P5-2, S1-1, P3-3, S2-1, P5-3, S3-2, and S2-2) did not show antibacterial activities in the streak test. These results indicate that living cells of the fish isolates inhibited the growth of reference strains.
[0112] Slides incorporated with extracts, living cells, and controls were deployed into the sea to determine if the secondary metabolites and living cells were active in situ against bacteria and eukaryotic organisms in the sea. Crude extracts (20 μl/slide) or washed cells (105 to 107 cells/ml) were incorporated into gels (Phytagel 3.26%) (Henrikson and Pawlik, 1995) on sterile, chemically cleaned glass slide. Crude extracts were obtained in the same manner as for antimicrobial experiments with the exception of using distilled water at pH 8.0 for the re-extraction, instead of EDTA buffer. Control slides were covered with Phytagel prepared with sterile extract controls. Treatments and controls were done in triplicates.
[0113] The gel covered ...

Problems solved by technology

Bacteria are unable to develop resistance against a s...

Method used

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  • Antagonistic properties of reef fish microflora
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Examples

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example 1

Sampling of Coral Reef Fish

[0096] The reef fish, Sparisoma ninidae (Parrotfish) and Lujanus purpureus (Red Snapper) were caught above a coral reef in True Blue Bay, Grenada at N=11°059.908 and W=061°46.282 in accordance with a Global Positioning System (GPS) (GEKO 201 Garmin Taiwan). Fish were either trapped in a fishpot or shot with a spear gun. The fishpot used to trap the fish was made of 0.5 inch galvanized square mesh, 36 inches long, 16 inches wide and high. The cornucol hole (horn shaped) on one side of the fishpot was 7 inches in diameter on the outer surface and it tapered inside the fishpot to a 5 inch diameter. The fishpot was located approximately 10 feet deep, next to a coral reef, approximately 150 feet away from shore. At the surface after caught, each fish was washed twice with autoclaved artificial seawater to remove any loosely associated microbes and then immediately placed in a sterile plastic bag on ice. The fish was then transported back to the laboratory.

example 2

Isolation of Pure Cultures from the Epithelial Mucosal Surfaces of Coral Reef Fish

[0097] Normal fish microflora was collected from the mucus surface of the fish with a sterile cotton swab and plated on Artificial Sea Water Agar (ASWA) medium. ASWA medium was used to mimic fish mucus. Artificial Sea Water Agar (ASWA) contained (g / l) of solution: NaCl 21.10, KCl 0.58, CaCl2×2H2O 1.20, MgCl2×6H2O 4.73, NaHCO3 0.08, MgSO4×7H2O 2.63, yeast extract 10.00, malt extract 4.00, glucose 4.00, agar 15.00. Solution was adjusted to pH 7.5-8.0, autoclaved at 121° C. for 20 min, and poured into sterile Petri dishes. Artifical Sea Water (ASW) liquid broth was prepared as above except the 15.00 g of agar was omitted.

[0098] Plates were incubated at 29° C. for 48 hours. Separate colonies were picked, inoculated and grown in the same liquid medium and cultured under the same temperature for 48 hours. Then the cultures were plated again on the solid ASWA. Gram staining was done to ensure that cultures ...

example 3

Morphological and Physiological Characterization of Isolates

[0101] Morphology of the cells from the isolates was studied after Gram-staining using phase contrast microscopy. Blood agar (Anon, 1953), Mannitol salts agar (Anon, 1953), Mac Conkey Agar (Anon, 1953), and Cystein tryptic agar (Anon, 1953) were used to test type of hemolysis, acid production from mannitol, resistance to bile salts, and oxidation / fermentation of glucose, respectively. Catalase reaction and oxidase reaction were performed using traditional techniques known in the art.

[0102] Twelve of the fifteen parrot fish isolates were gram positive, eight were cocoidal, and ten were pigmented. Three of the five snapper isolates were gram positive, four were cocoidal, and four were pigmented. The majority of the isolates were gram positive and pigmented (Table 2). Most of the isolated organisms were aerobic, heterotrophic, halotolerant, and mesophilic (Table 3). All of them grew best at 28° C. and salinity of 40 ppt, how...

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Abstract

The present invention provides methods for preventing biofilm formation on a surface. The present invention also relates to anti-biofilm forming agents, to methods of producing and using them, and to anti-fouling coatings produced therefrom.

Description

RELATED APPLICATIONS [0001] This application is a continuation-in-part of PCT / US05 / 015063, filed on May 2, 2005, which claims priority to U.S. provisional patent application Serial No. 60 / 566,600 filed Apr. 30, 2004 which are herein incorporated by reference in their entirety.BACKGROUND OF THE INVENTION [0002] Microbial biofilms cause systemic infections in humans and costly marine and industrial related damage and inefficiency. They cost billions of dollars yearly in equipment damage, product contamination, energy losses and medical infections. All living and non-living marine surfaces are potential sites for microbial biofilm formation. In the human body biofilms can be associated with tissues (e.g., inner ears, teeth, gums, lungs, heart valves and the urogenital tract) and on indwelling medical devices (e.g., contact lenses, central venous catheters and needleless connectors, endotracheal tubes, intrauterine devices, mechanical heart valves, pacemakers, peritoneal dialysis cathet...

Claims

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Application Information

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IPC IPC(8): A61K39/085A61K39/02C12N1/20A01N63/20H05B6/80
CPCA01N63/00A01N63/02C09D5/14C09D5/1687C12R1/01C09D5/1606A61P31/00A61P31/04A01N63/20C12R2001/01C12N1/205A01N63/10
Inventor BRUNO, CYNTHIA K.
Owner AEQUOR INC
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