Novalia Rachmawati
Research Center for Marine and Fisheries Product Processing and Biotechnology

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Skrining dan Identifikasi Bakteri Pembentuk Histamin yang Diisolasi dari Tuna, Tongkol, dan Cakalang Segar di Wilayah Jawa Barat, DKI Jakarta, dan Banten Novalia Rachmawati; Radestya Triwibowo
Jurnal Pascapanen dan Bioteknologi Kelautan dan Perikanan Vol 16, No 2 (2021): Desember 2021
Publisher : Balai Besar Riset Pengolahan Produk dan Bioteknologi Kelautan dan Perikanan

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.15578/jpbkp.v16i2.771

Abstract

Tuna, tongkol, cakalang (TTC) merupakan komoditas perikanan bernilai ekonomis penting yang disukai oleh banyak konsumen di Indonesia. Namun demikian, distribusi, penanganan, dan pengolahan komoditas ini masih banyak mengalami kendala, di antaranya kontaminasi bakteri pembentuk histamin (BPH) yang dapat menyebabkan akumulasi histamin dan menimbulkan kerugian kesehatan pada konsumen. Penelitian ini bertujuan untuk mengidentifikasi BPH dari komoditas TTC segar yang dijual di pasar domestik, mengevaluasi profilnya, serta mengkarakterisasi kemampuan BPH tersebut dalam menghasilkan histamin. Sebanyak 93 sampel TTC diperoleh dari TPI, pasar tradisional, dan pasar modern di wilayah Jawa Barat, DKI Jakarta, dan Banten. Dari 318 isolat presumtif BPH yang ditemukan, sebanyak 59 isolat (19%) terkonfirmasi positif gen hdc dan di antaranya sebanyak 43 isolat dikategorikan sebagai BPH prolifik. Hasil sekuensing 16S rDNA menunjukkan sebanyak 30 dari 43 isolat BPH prolifik (69,8%) adalah Morganella morganii. Selain M. morganii, isolat lain yang ditemukan dari semua jenis ikan yang diamati adalah Photobacterium damselae (6,9%), keduanya merupakan BPH mesofilik. Isolat mesofilik lain yang teridentifikasi dari sampel TTC berasal dari genus Klebsiella (4,7%), Proteus (4,7%), Raoultella (4,7%), Shewanella (2,3%), dan Vibrio (6,9%). Keberadaan BPH prolifik ini mengindikasikan adanya potensi akumulasi histamin pada produk akhir TTC apabila dalam penanganan dan pengolahannya tidak menerapkan sistem rantai dingin dengan benar.Title: Screening and Identification of Histamine Producing Bacteria Isolated from Fresh Tuna and Tuna-like from West Java, DKI Jakarta, and Banten AreasTuna and tuna-like fish are economically important and popular amongst Indonesian consumers. However, the distribution, handling, and processing of these commodities are still facing many problems, including contamination of histamine producing bacteria (HPB) which may lead to histamine accumulation and cause human adverse health effects. This study aimed to identify HPB from fresh tuna and tuna-like fish sold in domestic markets in Indonesia, evaluate their profile, and characterize their ability to produce histamine. A number of 93 fish samples were obtained from fish landing, traditional and modern fish markets in West Java, DKI Jakarta, and Banten. Of 318 presumptive HPB identified from the sample, 59 isolates (19%) were confirmed as hdc-gene positive with 43 isolates were categorized as prolific HPB. Bacterial identification with 16S rDNA sequencing identified 30 out of 43 (69.8%) prolific HPB as Morganella morganii. Besides M. morganii, another mesophilic HPB identified from all different type of fish was Photobacterium damselae (6.9%), while the remaining mesophilic HPB were identified from genus Klebsiella (4.7%), Proteus (4.7%), Raoultella (4.7%), Shewanella (2.3%), and Vibrio (6.9%). The presence of prolific HPB in the samples suggested that histamine accumulation in the final product is possible if cold-chain system is not properly applied during fish handling and processing.
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.