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EARLY DETECTION OF VIBRIOSIS ON POND WATER AND SEDIMENT

Ince Ayu K.Kadriah*, Nurhidayah
Research Institute for Coastal Aquaculture
Jl. Makmur Dg. Sitakka No. 129. Maros 90511, Sul-Sel
E-mail: inceayu@gmail.com

Outbreaks of Luminous Vibriosis caused by pathogenic Vibrio is a major constraint in shrimp production with potentially catastrophic impact.  The process of identification and detection using agar spread methods that can distinguish only the bacteria by phenotypic traits that are difficult to distinguish between pathogenic and non pathogenic bacteria.  Early detection of Vibriosis will provide accurate information about the contamination of Vibrio pathogens in aquaculture media before the population reaches a dangerous amount.  Molecular detection process having problems at the genome extraction stage.  Genomic extraction of Vibrio pathogen in ponds water and sediment require specific methods in order to extract the genome in very small amounts. In the sediment samples there are many other components that may interfere with the genome extraction process.  This study aims to determine the ability of molecular early detection methods for detecting vibriosis directly from pond water samples and sediment.

In this research we used a volume of 250ml Erlenmeyer.   Pond water that has been sterilized by autoclave put into erlenmeyer as 100mL. The concentration of pathogenic Vibrio harveyi in the water is set at 102, 103, 104, 105, 106, and 107 CFU/mL. Sampling is done by filtering 100mL of water that have been infected with the pathogenic Vibrio.  The extraction process uses Kit Power Water Mo Bio. Sediment ponds dried first before being used to dispose of toxic materials in the sediment.  For pond sediment research we used 250 mL Erlenmeyer filled pond sediment as much as 30g and plus seawater salinity 15ppt as much as 20ml.  Ponds sediment sterilized by autoclave at a temperature of 1210C at 1 atm pressure for 15 minutes. Culture pathogenic V. harveyi inserted into a sediment pond with concentration set of respectively 102, 103, 104, 105, 106, and 107 CFU/mL.  Each treatment was repeated three times. Sampling is done on days 2, 6 and 9. The genome extraction process from the sediment pond is done using Kit Power Soil Mo Bio. The result show that genome of Vibrio pathogens are not detected in water and sediment samples in all treatments. But PCR test showed that Vibrio pathogens can be detected although its genome is not looked at the results of electrophoresis.




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