Sains Malaysiana 53(1)(2024): 99-110
http://doi.org/10.17576/jsm-2024-5301-08
In-vitro Inhibitory Activities of Potential Probiotic Isolated from Pangasius
nasutus against Aeromonas hydrophila and Streptococcus agalactiae
(Aktiviti Perencatan In-vitro Probiotik Berpotensi Diasingkan daripada Pangasius nasutus terhadap Aeromonas
hydrophila dan Streptococcus agalactiae)
SHAFIQ JOHAR1,
PUVANESWARI PUVANASUNDRAM1,2, CLEMENT ROY DE CRUZ1,3,
CHOU MIN CHONG1,2, MD YASIN INA-SALWANY1,2, KENG CHIN LIM1,
NOORDIYANA MAT NOORDIN4 AND MURNI KARIM1,3,*
1Department
of Aquaculture, Faculty of Agriculture, Universiti Putra Malaysia, 43400 UPM
Serdang, Selangor, Malaysia
2Aquatic Animal Health and Therapeutics Laboratory,
Institute of Bioscience, Universiti Putra Malaysia, 43400 UPM Serdang,
Selangor, Malaysia
3Laboratory
of Sustainable Aquaculture, International Institute of Aquaculture and Aquatic
Sciences, Universiti Putra Malaysia, 71050 Port Dickson, Negeri Sembilan,
Malaysia
4Faculty of Fisheries and Food Science, Universiti
Malaysia Terengganu, 21030 Kuala Nerus, Terengganu, Malaysia
Diserahkan:
27 Oktober 2023/Diterima: 28 Disember 2023
Abstract
In aquaculture, using probiotics
is crucial for strengthening the immune system and encouraging the growth and
survival of many aquatic organisms, including the Pangasius species.
This approach is particularly significant given the impact of bacterial diseases
on Pangasius survival. This study aimed to assess the effectiveness of
probiotics isolated from Pangasius nasutus as alternatives to
antibiotics for combating infections caused by Aeromonas hydrophila and Streptococcus
agalactiae. Potential bacteria were isolated from the intestine and stomach
of healthy P. nasutus. Seventy probiotic strains were successfully
isolated and further screened using A. hydrophila and S. agalactiae as pathogens in an in vitro disc diffusion assay. Preliminary screenings
indicated that five probiotic strains inhibited the growth of A. hydrophila. Stomach-derived strain S1 and intestine-derived strain L1 suppressed A.
hydrophila growth with inhibition zones of 10.5±1 mm and 8.5±1 mm,
respectively. Likewise, strains L2, L8, and L12 from the intestine showed
inhibitory zones of 6.0±1 mm, 6.5±1 mm, and 6.0±1 mm, respectively. Of these,
only L12 inhibited the growth of S. agalactiae with a clear zone of
5.0±1 mm. In the elimination of pathogenic strains, potential strains S1 and L1
did not grow on the Aeromonas isolation medium. Co-culture assays
demonstrated that both potential strains significantly inhibited Aeromonas
hydrophila growth at concentrations of 106 and 108 CFU
mL-1 over 48- and 96-hour periods, respectively. The potential
bacterial strains were identified using 16s rRNA gene sequencing and were
classified as follows: S1 - Lactococcus lactis, L1 - Weissella
confusa, L2 - Cosenzaea myxofaciens, L8 - Lactococcus garvieae, and L12 - Plesiomonas shigelloides. Strain S1 L. lactis and
strain L1 W. confusa are suggested for further evaluation and acquired
additional research to fully elucidate their mechanisms and potential as
probiotics.
Keywords: Antagonistic activity; in
vitro screening; Pangasius species; pathogens; potential probiotics
Abstrak
Dalam
akuakultur, penggunaan probiotik adalah penting untuk menguatkan sistem
keimunan dan menggalakkan pertumbuhan dan kemandirian hidup pelbagai organisma
akuatik, termasuk spesies Pangasius. Pendekatan ini amat penting
memandangkan kesan buruk penyakit bakteria terhadap kelangsungan hidup spesies Pangasius.
Kajian ini bertujuan untuk menilai keberkesanan probiotik yang dipencilkan
daripada Pangasius nasutus sebagai alternatif kepada antibiotik untuk
melawan jangkitan penyakit yang disebabkan oleh Aeromonas hydrophila dan Streptococcus agalactiae. Bakteria berpotensi sebagai probiotik telah
diasingkan daripada organ usus dan perut P. nasutus yang sihat. Tujuh
puluh strain probiotik berjaya diasingkan dan seterusnya disaring dengan
menggunakan A. hydrophila dan S. agalactiae sebagai patogen dalam
ujian penyebaran cakera in vitro. Pemeriksaan awal menunjukkan bahawa
lima strain probiotik dapat merencat pertumbuhan A. hydrophila. Strain S1 yang dipencilkan daripada perut dan strain L1 daripada usus dapat merencat
pertumbuhan A. hydrophila dengan zon perencatan masing-masing sebanyak
10.5±1 mm dan 8.5±1 mm. Begitu juga, strain L2, L8 dan L12 daripada usus
menunjukkan zon perencatan ke atas A. hydrophila masing-masing sebanyak
6.0±1 mm, 6.5±1 mm dan 6.0±1 mm. Manakala, hanya L12 merencat pertumbuhan S.
agalactiae dengan zon yang jelas 5.0±1 mm. Dalam pengasingan strain patogen, S1 dan L1 tidak hidup di atas medium pengasingan Aeromonas.
Ujian kultur bersama menunjukkan bahawa kedua-dua SI dan LI dengan ketara
merencat pertumbuhan A. hydrophila pada kepekatan 106 dan 108 CFU mL-1 dalam tempoh 48 dan 96 jam. Semua strain bakteria
dikenal pasti menggunakan penjujukan gen 16s rRNA dan dikelaskan seperti
berikut: S1 - Lactococcus lactis, L1 - Weissella confusa, L2 - Cosenzaea
myxofaciens, L8 - Lactococcus garvieae dan L12 - Plesiomonas
shigelloides. Strain SI L. lactis dan L1 W. confusa dicadangkan untuk penilaian lanjut dan memerlukan penyelidikan tambahan untuk
menjelaskan sepenuhnya mekanisme dan potensi sebagai probiotik.
Kata kunci: Aktiviti antagonis; kajian in vitro; patogen; probiotik berpotensi;
spesies Pangasius
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*Pengarang untuk surat-menyurat; email:
murnimarlina@upm.edu.my
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