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γ-Aminobutyric acid production by selected lactic acid bacteria isolate of an Indonesian indigenous fermented buffalo milk (dadih) origin

Harnentis H, Nurmiati N, Marlida Y, Adzitey F, Huda N

Vet World, 2019 Aug;12(8):1352-1357.
PMID: 31641319 DOI: 10.14202/vetworld.2019.1352-1357

Aim: This study aimed at optimizing γ-aminobutyric acid (GABA) production using lactic acid bacteria (LAB) of an Indonesian indigenous fermented buffalo milk (dadih) origin. This study utilized LAB previously cultured from dadih that has the ability to produce GABA.
Materials and Methods: The study started with the identification of selected LAB by 16S rRNA, followed by optimization of GABA production by culture conditions using different initial pH, temperature, glutamate concentration, incubation time, carbon, and nitrogen sources. 16S rRNA polymerase chain reaction and analysis by phylogenetic were used to identify Lactobacillus plantarum (coded as N5) responsible for the production of GABA.
Results: GABA production by high-performance liquid chromatography was highest at pH of 5.5, temperature of 36°C, glutamate concentration of 500 mM, and incubation time of 84 h. Peptone and glucose served as the nitrogen and carbon sources, respectively, whereas GABA was produced at optimum fermentation condition of 211.169 mM.
Conclusion: Production of GABA by L. plantarum N5 was influenced by initial pH of 5.5, glutamic acid concentration, nitrogen source, glucose as carbon source, and incubation temperature and time.
Fulltext Molecular identification and phylogenetic analysis of GABA-producing lactic acid bacteria isolated from indigenous dadih of West Sumatera, Indonesia

Anggraini L, Marlida Y, Wizna W, Jamsari J, Mirzah M, Adzitey F, et al.

F1000Res, 2018 10 19;7:1663.
PMID: 32201563 DOI: 10.12688/f1000research.16224.3

Background: Dadih (fermented buffalo milk) is a traditional Indonesian food originating from West Sumatra province. The fermentation process is carried out by lactic acid bacteria (LAB), which are naturally present in buffalo milk. Lactic acid bacteria have been reported as one of potential producers of γ-aminobutyric acid (GABA). GABA acts as a neurotransmitter inhibitor of the central nervous system. Methods: In this study, molecular identification and phylogenetic analysis of GABA producing LAB isolated from indigenous dadih of West Sumatera were determined. Identification of the GABA-producing LAB DS15 was based on conventional polymerase chain reaction. 16S rRNA gene sequence analysis was used to identify LAB DS15. Results: PCR of the 16S rRNA gene sequence of LAB DS15 gave an approximately 1400 bp amplicon. Phylogenetic analysis showed that LAB DS15 was Pediococcusacidilactici, with high similarity of 99% at 100% query coverage to Pediococcusacidilactici strain DSM 20284. Conclusions: It can be concluded that GABA producing LAB isolated from indigenous dadih was Pediococcus acidilactici.
Fulltext Novel probiotic lactic acid bacteria isolated from indigenous fermented foods from West Sumatera, Indonesia

Harnentis H, Marlida Y, Nur YS, Wizna W, Santi MA, Septiani N, et al.

Vet World, 2020 Sep;13(9):1922-1927.
PMID: 33132606 DOI: 10.14202/vetworld.2020.1922-1927

Background and Aim: Probiotics play an important role in maintaining a healthy gut and consequently promote good health. This study aimed to find novel probiotic lactic acid bacteria (LAB) from indigenous fermented foods of West Sumatera, Indonesia.
Materials and Methods: This study utilized 10 LAB previously isolated from fermented buffalo milk (dadih), fermented fish (budu), and fermented cassava (tape) which have the ability to produce gamma-aminobutyric acid. The study commenced with the screening of LAB for certain properties, such as resistance to acid and bile salts, adhesion to mucosal surface, and antagonism against enteric pathogens (Escherichia coli, Salmonella Enteritidis, and Staphylococcus aureus). The promising isolates were identified through biochemical and gram staining methods.
Results: All isolates in this study were potential novel probiotics. They survived at a pH level of 2.5 for 3 h (55.27-98.18%) and 6 h (50.98-84.91%). Survival in bile at a concentration of 0.3% was 39.90-58.61% and the survival rate was 28.38-52.11% at a concentration of 0.5%. The inhibitory diameter ranged from 8.75 to 11.54 mm for E. coli, 7.02 to 13.42 mm for S. aureus, and 12.49 to 19.00 mm for S. Enteritidis. All the isolates (84.5-92%) exhibited the ability to adhere to mucosal surfaces. This study revealed that all the isolates were potential probiotics but N16 proved to be superior because it was viable at a pH level of 2 (84.91%) and it had a good survival rate in bile salts assay (55.07%). This isolate was identified as Lactobacillus spp., Gram-positive bacilli bacteria, and tested negative in both the catalase and oxidase tests.
Conclusion: All the isolates in this study may be used as probiotics, with isolate N16 (Lactobacillus spp.) as the most promising novel probiotic for poultry applications based on its ability to inhibit pathogenic bacteria.
The potential of lactic acid bacteria isolated from ikan budu (fermented fish) to inhibit the growth of pathogenic fungi and detoxify aflatoxin B1

Marlida Y, Nurmiati N, Husmaini H, Huda N, Anggraini L, Ardani LR

Vet World, 2023;16(7):1373-1379.
PMID: 37621548 DOI: 10.14202/vetworld.2023.1373-1379

BACKGROUND AND AIM: Market demand for safe feed and food supply and consumer preferences for safe and healthy products are increasing. Control measures to counter threats to the feed supply need to be implemented as early as possible to prevent economic losses. Mycotoxins produced by certain groups of fungi are a problem that can disrupt the feed supply or pose a threat to the health of animals and humans. Biological control to detoxify contaminated feed ingredients can be carried out on a large scale economically. For example, lactic acid bacteria (LAB) can act as biological agents for eliminating mycotoxins. This study aimed to clarify the value of screening LAB to inhibit Aspergillus flavus growth and detoxify aflatoxin B1 (AFB1).
MATERIALS AND METHODS: In this study, using a completely randomized design with three replications, five isolates of LAB (LA.1, LA.6, LA.8, LA.12, and LA.22) along with their supernatants were tested qualitatively and quantitatively for their ability to counter mycotoxins using A. flavus and corn kernels. The isolates with the best activity were identified by sequencing 16S rDNA.
RESULTS: The results showed that the five LAB isolates can inhibit the growth of A. flavus and detoxify AFB1. Among these isolates, LA.12 showed the best performance, followed by LA.22, LA.8, LA.6, and then LA.1. The sequencing results confirmed that LA.12 was Lactobacillus harbinensis strain 487.
CONCLUSION: All of the isolates in this study have the potential as biological agents for detoxifying AFB1, with isolate LA.12 appearing to be the most promising biodetoxification agent for feed (AFB1 in corn) based on its ability to inhibit pathogenic fungi.
Fulltext Metagenomic analysis and biodiversity of bacteria in traditional fermented fish or Budu from West Sumatera, Indonesia

Marlida Y, Susalam MK, Harnentis H, Jamsari J, Huda N, Noordin WNM, et al.

J Adv Vet Anim Res, 2023 Dec;10(4):801-808.
PMID: 38370893 DOI: 10.5455/javar.2023.j736

OBJECTIVE: This research aims to investigate the microbial diversity of Budu prepared from fresh and frozen fish from the Pariaman and Pasaman districts in West Sumatra Province, Indonesia, as well as provide basic information about Budu quality.
MATERIALS AND METHODS: To obtain the bacterial microbial composition, deoxyribonucleic acid extraction was carried out using amplicon-sequencing of the 16S-rRNA gene in the V3-V4 region from two types of Budu and carried out in duplicate.
RESULTS: Budu prepared with fresh (Pariaman) or frozen (Pasaman) fish was dominated by Firmicutes (78.455%-92.37%) and Proteobacteria (6.477%-7.23%) phyla. The total microbial species in Budu from Pariaman were higher (227 species) than in Pasaman (153 species). The bacterial species found are Lentibacillus kimchi (1.878%-2.21%), Staphylococcus cohnii (0.597%-0.70%), Peptostreptococcus russeli (0.00%-0.002%), Clostridium disporicum (0.073%-0.09%), Clostridium novyi (0.00%-0.01%), Nioella sediminis (0.00%-0.001%), and Shewanella baltica (0.00%-0.003%). Lentibacillus kimchi, S. cohnii, and C. disporicum are found in both Budu. Nioella sediminis and S. baltica are found in Budu Pariaman. Peptostreptococcus russeli and C. novyi were found in Budu Pasaman.
CONCLUSION: Metagenomic analysis of Budu from different fish, Pariaman (fresh fish) and Pasaman (frozen fish) showed that the biodiversity of bacteria was barely different. Both Budu found lactic acid bacteria from the Enterococcaceae family, genus Vagococcus, and pathogenic bacteria, such as S. cohnii, P. russeli, C. disporicum, and S. baltica. The discovery of various species of pathogenic bacteria indicates that development is still needed in the Budu production process to improve Budu quality.