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Black Sea Luminous Bacteria and their Applications

    Andrew Katsev

Keywords: luminous bacteria, luciferase, biotest

Bacteria origins and properties

In 1998 a private initiative project for studying the features of luminous bacteria from the Black Sea and the Azov Sea in the Crimean region (Ukraine) was begun. From a series of samples (sea-water and sea-animals) obtained mainly from the coastal zone of Black Sea in the summer seasons (1998 - 2000) 10 strains of luminous bacteria were isolated and identified, as:
  • Photobacterium phosphoreum - 3 strains (isolated from Squalus acanthias, shallow fish)
  • Vibrio fischeri, V.logei - 5 strains (from Squalus acanthias, Rapana
    thomasiana, shallow fish, shrimp)
  • Photobacterium sp. - 1 strain (from a mussel)
  • Vibrio sp. - 1 strain (from a shrimp)
The last two isolates are not complete identified yet.

The morphological, physiological and biochemical properties of these strains, and also kinetic properties of their luciferases are investigated.
The assessment of bioluminescence sensitivity of these strains to some toxicants (ions of heavy metals, phenol, potassium bichromate, sodium dodecyl sulphate) in acute and chronic toxicity tests in comparison with Ph. phosphoreum (Cohn) Ford (it's an official strain recommended to use for toxicity assessment in Ukraine) has shown their applicability for toxicity biotesting. The strains are planned to use for study an ecological situation in Crimea.

Medical application of intact luminous bacteria

Lipoplysaccharide assay
Bioluminescent technique is used for determination of bacterial lipoplysaccharide upon reaction with polymixin. Bacterial luminescence level is closely depended of the free antibiotic amounts. It is shown that the reaction between lipoplysaccharide and polymixin B results in reduction of antibiotic action and restoration the bioluminescence level. This effect was used as a method for quantitative lipoplysaccharide analyses. Obtained curves make it possible to determine lipoplysaccharide from E.coli K30 and S. typhia at a range of 0.5 - 5 mg/ml.

Albumin binding capacity determination
The binding of cationic surface-active antiseptics by seralbumin is studied with the usage of the Black Sea luminous bacteria Photobacterium phosphoreum. It is shown that the binding of cationic antiseptics with seralbumin resulted in petering toxic and antimicrobial properties of aethonium, miramistinum and decamethoxinum, but not chlorhexidinum. At the presence of
albuminum the activity of chlorhexidinum was reduced only on 45 - 48%. As defined, the albumin binding capacity to miramistinum was about - 7.0 - 8.7%; aethonium - 8.2 - 10.0% and decamethoxinum - 2.4 - 3.0%.

Monitoring of psychopharmacological substances
Bioluminescent technique was used for bioassay of ten psychopharmacological substances at the presence of physiological liquids (serum, saliva and urine). The group of phenothiazine derivatives was more active in the bacterial and luciferase tests. It is shown that the most perspective environments for the drugs biotesting are saliva and urine. At the presence of serum the effect of the substances decreases because of their links with proteins (serum albumin).

Diagnosis of dental diseases
Isolated luminous strains have been used for mixed saliva biotesting.
Experimental conditions were optimized. The correlation between the level of bacterial luminescence in the presence of saliva and intensity of dental disease was found out. In the case of caries and periodontal diseases the saliva effect on bioluminescence was on 30 40% less than in healthy mouth.

Submitted: 9-Apr-2001

The contents of this page are the sole responsibility of the person named below.

Andrew Katsev |
Crimean Medical University |
Boulevard Lenina 5/7 |
Simferopol, Crimea Ukraine |
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