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Sains Malaysiana 54(12)(2025): 2847-2858
http://doi.org/10.17576/jsm-2025-5412-04
Screening
and Preliminary Characterization of Potential
Probiotic Lactic Acid Bacteria from Vegetable
Wastes
NUR
HIDAYAHANUM HAMID1,*, MOHD FAKHRULDDIN
ISMAIL1, HASSAN MOHD DAUD2, PRAPANSAK SRISAPOOME3, MURNI MARLINA ABD KARIM1,5, AHMAD ZAHARIN ARIS1, RUHIL
HAYATI HAMDAN4 & SITI FAIRUS
MOHAMED YUSOF5
1International Institute of Aquaculture
and Aquatic Sciences (I-AQUAS), Universiti Putra Malaysia, 70150 Port Dickson, Negeri Sembilan, Malaysia
2Aquatic Animal Health Unit,
Faculty of Veterinary Medicine, Universiti Putra
Malaysia, 43400 UPM Serdang, Selangor, Malaysia
3Laboratory of Aquatic
Animal Health Management, Faculty of Fisheries, Kasetsart University, Bangkok 10900, Thailand
4Faculty of Veterinary Medicine, Department of
Paraclinical Studies, Universiti Malaysia Kelantan,
16100 Kota Bharu, Kelantan, Malaysia
5Department of Aquaculture, Faculty of Agriculture, Universiti Putra Malaysia, 43400 UPM Serdang, Selangor,
Malaysia
Received: 20 February 2025/Accepted: 10 December 2025
Abstract
Southeast Asia generates
over 8 million tons of vegetable waste annually, yet the microbial isolations
from these wastes are rarely associated with potent probiotics. This study
aimed to screen lactic acid bacteria (LAB) from fermented and non-fermented
vegetable wastes for probiotic potential. LAB were isolated using serial dilutions on MRS agar supplemented with 1% CaCO3, yielding 17 isolates. Two isolates, FCU21 and FBS34, demonstrated notable probiotic
potential by exhibiting antibacterial activity against Aeromonas hydrophila (6.3 ± 0.08 mm and 6.2 ± 0.04 mm) and Streptococcus agalactiae (5.6 ± 0.04 mm and 10.4 ±
0.28 mm). Initial identification
based on morphological, phenotypic and biochemical characteristics showed Gram-positive,
catalase-negative, non-spore-forming cocci. Genotypic analysis using 16S rRNA
sequencing confirmed both isolates as Enterococcus hirae,
with 99% similarity for FCU21 (from fermented cucumber) and 100% for FBS34
(from fermented mung bean sprouts), designated as Enterococcus hirae UPM01 and UPM02, respectively. Whole-genome
shotgun sequencing showed distinct genomic features, including variations in base
pairs, contigs and coding sequences. These findings highlight the promising
potential of Enterococcus hirae strains as
probiotics for aquaculture, warranting further investigation into their
probiotic properties and applications.
Keywords: Enterococcus hirae; lactic acid bacteria; probiotic; vegetable
waste
Abstrak
Asia Tenggara menghasilkan lebih daripada 8 juta tan sisa sayuran setiap
tahun, namun mikroorganisma yang dipencilkan daripada sisa ini jarang dikaitkan
dengan probiotik yang berpotensi tinggi. Penyelidikan ini bertujuan untuk
menilai bakteria asid laktik (LAB) daripada sisa sayuran yang ditapai dan tidak
ditapai bagi penilaian potensi probiotik. LAB diasingkan menggunakan pencairan bersiri pada agar MRS yang ditambah dengan 1% CaCO3, menghasilkan 17 pencilan. Dua pencilan, FCU21 dan
FBS34 menunjukkan potensi probiotik yang lebih baik dengan aktiviti antibakteria terhadap Aeromonas hydrophila (6.3 ± 0.08 mm dan
6.2 ± 0.04 mm) dan Streptococcus agalactiae (5.6 ± 0.04 mm dan 10.4 ± 0.28 mm). Pengenalpastian awal berdasarkan ciri morfologi, fenotip dan biokimia menunjukkan pencilan tersebut adalah kokus Gram-positif, katalase-negatif dan tidak membentuk spora. Analisis genotip menggunakan penjujukan 16s rRNA mengesahkan kedua-dua pencilan sebagai Enterococcus hirae dengan persamaan 99% untuk FCU (daripada timun yang ditapai) dan 100% untuk FBS4 (daripada taugeh kacang hijau yang ditapai), yang kemudiannya dinamakan sebagai Enterococcus hirae UPM01 dan UPM02. Penjujukan seluruh genom mendedahkan ciri genom yang berbeza, termasuk variasi dalam pasangan bes, kontig dan jujukan pengekodan. Penemuan ini menyerlahkan potensi besar strain Enterococcus hirae sebagai probiotik untuk akuakultur yang memerlukan kajian lanjut mengenai sifat dan aplikasi probiotiknya.
Kata kunci: Bacteria asid laktik; Enterococcus hirae; probiotik; sisa sayuran
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*Corresponding author; email: hidayahanum@upm.edu.my
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