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All Journal Squalen Bulletin of Marine and Fisheries Postharvest and Biotechnology
Dwiyitno Dwiyitno
Research Center For Marine and Fisheries Product Processing and Biotechnology
Agriculture, Biological Sciences & Forestry Biochemistry, Genetics & Molecular Biology Environmental Science Immunology & microbiology

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Rapid and Simultaneous Detection of Vibrio parahaemolyticus, Salmonella spp. and Escherichia coli in Fish by Multiplex PCR Radestya Triwibowo; Novalia Rachmawati; Dwiyitno Dwiyitno
Squalen, Buletin Pascapanen dan Bioteknologi Kelautan dan Perikanan Vol 15, No 2 (2020): August 2020
Publisher : Research and Development Center for Marine and Fisheries Product Processing and Biotechnol

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.15578/squalen.v15i2.444

Abstract

Pathogenic bacteria are commonly found as natural contaminants in seafood and fish products. Globally, several countries have been imposing strict regulations on the maximum levels of pathogens and consequently require microbial testing of pathogens before the products can be marketed. A culture-based method with biochemical assay has been widely used to detect pathogenic bacteria in food, despite its long and extensive process. Meanwhile, the alternative molecular-based method to overcome this problem, cannot differentiate between viable and nonviable cells, which may lead to underestimation. This study aimed to develop a multiplex PCR (mPCR) method as a confirmatory assay for the culture-based method to detect pathogens in fish products simultaneously. This method applied a pre-enrichment step to ensure the growth of low-level pathogens and the injured cells in the sample. The target genes were ToxR, InvA, and UidA for Vibrio parahaemolyticus, Salmonella spp. and Escherichia coli, respectively. This assay also amplified the 16S rDNA gene of bacteria as an internal control for the PCR reaction. By implementing liquid-based DNA extraction during analysis, the developed-mPCR was comparable to detect the targeted bacteria in artificially-contaminated samples. The method was more sensitive in naturally-contaminated samples, where the number of E. coli, Salmonella spp. and V. parahaemolyticus detected were 28, 7, and 22, respectively. While the conventional method only detected 26, 5, and 19 of the respective pathogens. With a relatively shorter time and lower operation cost, the mPCR method is potential as an alternative for the culture-based method.
Seasonal and Spatial Water Pollution Status at The Rivers Estuaries in Cirebon Region Izhamil Hidayah; Hedi Indra Januar; Dwiyitno Dwiyitno; Nining Betawati Prihantini
Squalen, Buletin Pascapanen dan Bioteknologi Kelautan dan Perikanan Vol 16, No 2 (2021): August 2021
Publisher : Research and Development Center for Marine and Fisheries Product Processing and Biotechnol

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.15578/squalen.505

Abstract

Cirebon has 18 rivers that flow into the Java Sea. The Cirebon rivers play an important role in several sectors such as industrial, agriculture, household, and also aquaculture. The increasing anthropogenic activities, may have affected the health and quality of the Cirebon rivers and the surrounding area. This study aimed to determine the water pollution level of Cirebon rivers based on the Storage and Retrieval (STORET) and Pollution Index approaches. The study was conducted at seven different rivers located in the districts (kabupaten) and municipals (kotamadya) of Cirebon. The observation was performed during the end of the west monsoon (March 2019) as well as the beginning (July 2019) and the end of the east monsoon (November 2019). The STORET score of the Cirebon district rivers were between -10 and -20, which is categorized as lightly to moderately polluted. Similarly, most of the Cirebon municipal rivers were moderately contaminated throughout the year (-18 to -30). In contrast, the Pollution Index (PI) values of all observed rivers were between 1.0 to 5.0 which are categorized as lightly polluted.
Co-Authors Hedi Indra Januar Izhamil Hidayah Nining Betawati Prihantini Novalia Rachmawati Radestya Triwibowo