Identifikasi Bakteri Vibrio Parahaemolyticus Pemicu Penyakit Acute Hepatopancreatic Necrosis Disease (AHPND) Pada Udang Dengan Metode Spektroskopi Fluoresens

Salsabila, Vena (2023) Identifikasi Bakteri Vibrio Parahaemolyticus Pemicu Penyakit Acute Hepatopancreatic Necrosis Disease (AHPND) Pada Udang Dengan Metode Spektroskopi Fluoresens. Masters thesis, Institut Teknologi Sepuluh Nopember.

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Abstract

Penyakit acute hepatopancreatic necrosis disesase (AHPND) atau yang dikenal sebagai early mortality syndrome (EMS) merupakan penyakit yang menyerang udang disebabkan oleh bakteri Vibrio parahaemolyticus dengan tingkat kematian hingga 100%. Untuk mencegah penyakit AHPND, maka salah satu upaya paling penting adalah rapid test untuk identifikasi bakteri V. parahaemolyticus beserta konsentrasinya sebagai screening awal agar dapat mengontrol maupun memberikan penanganan dengan segera. Standar bakteri V. parahaemolyticus didapatkan melalui isolasi bakteri melalui tiga sumber berbeda dengan masing masing tiga kali pengulangan yaitu air bak benur, air bak maturasi, dan body benur. Metode yang digunakan yaitu kombinasi metode spread plate dan metode streaking. Sedangkan media yang digunakan adalah media chrom agar dan sea water complete (SWC) sebagai media cair. Bakteri V. parahaemolyticus ditandai dengan munculnya koloni bakteri berwarna mauve. Analisis fluoresens dilengkapi teknik synchronous fluorescence spectroscopy digunakan untuk mendapatkan panjang gelombang khas (λeks dan λem) V. parahaemolyticus dengan scan variasi interval Δλ 20-200 nm agar didapatkan panjang gelombang paling optimum. Melalui spektra dan contour plot, didapatkan λeks = 281 nm pada Δλ 70, sedangkan λem = 351 nm (λem = λeks +Δλ). Analisis real sampel secara kualitatif dan kuantitatif juga diaplikasikan menggunakan supernatan body benur yang telah melalui proses preparasi, dengan metode isolasi bakteri media chrom agar sebagai metode validasi. LOD metode fluoresens pada bakteri murni (kultivasi) V. parahaemolyticus pada sumber yang berbeda adalah 104–105 CFU/mL. Sedangkan LOD V. parahaemolyticus dalam benur udang sebesar 6,9 CFU/mL. Kombinasi metode fluoresens dengan Principal Component Analysis (PCA) sebagai multivariate analysis, dapat mengklasterisasi bakteri V. parahaemolyticus dengan bakteri pembanding meski dalam satu genus. Selain itu, melalui PCA juga dapat membedakan benur sehat dengan benur yang terpapar V. parahaemolyticus. Metode ini terbukti cepat, selektif, murah, dan tidak memerlukan preparasi rumit dibandingkan dengan metode deteksi bakteri lainnya 
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Acute hepatopancreatic necrosis disease (AHPND) or known as early mortality syndrome (EMS) is a disease that attacks shrimp caused by the bacterium Vibrio parahaemolyticus with a mortality rate of up to 100%. To prevent AHPND disease, one of the most important efforts is a rapid test to identify the presence of V. parahaemolyticus bacteria and their concentration as an initial screening so that they can control and provide immediate treatment. The standard of V. parahaemolyticus bacteria was obtained by isolating the bacteria from three different sources with three repetitions each, namely post-larvae tub water, maturation bath water, and post-larvae body by combining the spread plate method and the streaking method. The media used were Chrom agar media and seawater complete (SWC) as a liquid medium parahaemolyticus bacteria are characterized by the appearance of mauve-colored bacterial colonies. The fluorescence analysis is equipped with synchronous fluorescence spectroscopy techniques to obtain the typical wavelengths (λex and λem) of V. parahaemolyticus with scan interval variations of Δλ 20-200 nm in order to obtain the most optimum wavelength. Through spectra and contour plots, we get λeks = 281 nm at Δλ 70, while λem = 351 nm (λem = λeks +Δλ). Qualitative and quantitative analysis of the real sample was also applied using post-larvae body supernatants that had gone through the preparation process, with the bacterial isolation method in Chrom agar media as a validation method. The LOD of the fluorescence method on pure V. parahaemolyticus bacteria (cultivation) at different sources was 104–105 CFU/mL. Meanwhile, the LOD of V. parahaemolyticus in the body of post-larvae shrimp was 6,9 CFU/mL. The combination of the fluorescence method with Principal Component Analysis (PCA) as a multivariate analysis, can clustery V. parahaemolyticus bacteria with comparison bacteria even in the same genus. In addition, PCA can also differentiate between a healthy shrimp and an exposure V. rahaemolyticus. This method is proven to be fast, selective, inexpensive, and does not require complicated preparations compared to other bacterial detection methods

Item Type:	Thesis (Masters)
Uncontrolled Keywords:	V. parahaemolyticus, AHPND, fluoresens, synchronous fluorescence spectroscopy, V. parahaemolyticus, AHPND, fluorescence, synchronous fluorescence spectroscopy
Subjects:	Q Science > QD Chemistry > QD75.2 Chemistry, Analytic Q Science > QD Chemistry > QD96F56 Fluorescence spectroscopy R Medicine > RB Pathology S Agriculture > SH Aquaculture. Fisheries. Angling
Divisions:	Faculty of Science and Data Analytics (SCIENTICS) > Chemistry > 47101-(S2) Master Thesis
Depositing User:	Vena Salsabila
Date Deposited:	09 Feb 2023 07:39
Last Modified:	09 Feb 2023 07:39
URI:	http://repository.its.ac.id/id/eprint/96548

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