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MOLECULAR IDENTIFICATION AND ANTIBACTERIAL ANALYSIS OF LACTIC ACID BACTERIA FROM COCONUT WATER (COCOS NUCIFERA) AS A PROBIOTIC CANDIDATE

Open access
Suryani, Suryani, Taslim, Tuty, Darmana, Ija, Rahmawati, Rahmawati, Arikhman, Nova, Martinus, B.A, Chrisnawati, Chrisnawati, Syamsuwirman, Syamsuwirman·2023·RASAYAN Journal of Chemistry
DOI 10.31788/rjc.2023.1628239
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Abstract

Coconut water has been traditionally used as a fever medicine by the Minangkabau people in Indonesia since ancient times, but the chemical or probiotic components have not been widely studied. Therefore, this study aims to isolate and molecularly identify the probiotics found in coconut water, as well as to analyze their antimicrobial and antioxidant abilities. The Lactic acid bacteria (LAB) isolation was conducted by applying MRSA + CaCO3 with a concentration of 0.5%, and the molecular identification used the PCR (Polymerase Chain Reaction) method. Meanwhile, morphological and physiological identifications were also carried out with biochemical tests. The agar diffusion method was used to analyantimicrobialsial by measuring the diameter of the inhibition zone. The results showed that 97 isolates were lactic acid bacteria, while morphological, physiological, and biochemical tests showed Lactobacillus paracasei, Lactobacillus plantarum, Pediococcus sp 1, Pediococcus sp 2, Pediococcus sp 3, and Pediococcus sp 4, as well as two species of fungi such as Candida sp and Rizhopus. The molecular identification showed that Enterococcus faecalis strain 2358 can inhibit the growth of Staphylococcus aureus. Therefore, coconut water can be traditionally used to treat fever medicine because it contains probiotics and antimicrobial properties

AI snapshot

Written from the paper’s full text.

TL;DR

Fresh coconut water yielded 97 lactic acid bacteria isolates on MRSA + CaCO3, with morphological/biochemical tests identifying Lactobacillus and Pediococcus groups and molecular identification reporting Enterococcus faecalis strain 2358. The isolate inhibited four test bacteria in agar diffusion assays, supporting the traditional use of coconut water as a probiotic/antimicrobial remedy.

Key findings

  • A total of 97 LAB isolates were obtained from fresh coconut water; Table 1 reports 6 colonies at 10⁻⁵, 38 at 10⁻⁶, and 53 at 10⁻⁷.
  • Morphological and biochemical testing showed all isolates were Gram-positive; 49 were rod-shaped and the remainder coccus-shaped, with negative catalase and motility results. Table 2 lists 12 Lactobacillus paracasei, 37 Lactobacillus plantarum, 14 Pediococcus sp 1, 19 Pediococcus sp 2, 21 Pediococcus sp 3, and 14 Pediococcus sp 4.
  • Molecular identification by PCR and phylogenetic tree analysis identified Enterococcus faecalis strain 2358. In antimicrobial testing, average inhibition zones were 25 mm for E. coli, 25.3 mm for L. monocytogenes, 23.6 mm for S. typhi, and 24.3 mm for S. aureus; the text states the largest zones were against L. monocytogenes and E. coli and the smallest against S. typhi.

Methods

  • Fresh coconut water was diluted in sterile saline and plated by pour-plate on MRSA + CaCO3 selective medium, then incubated overnight at 37°C; colonies growing in clear “halo” zones were selected.
  • Isolates were characterized by Gram staining, catalase test, motility test, and colony morphology. DNA was extracted using a Chelex/GES method, amplified by PCR for 16S rDNA with primers 27F and 1492R, Sanger-sequenced using primers 518F, 800R, and 1492R on an ABI 3130 XL, and used to construct a phylogenetic tree.
  • Antimicrobial activity was analyzed by agar diffusion against Staphylococcus aureus, Escherichia coli, Listeria monocytogenes, and Salmonella typhi; test bacteria were standardized to McFarland 0.5, incubated overnight at 37°C, and inhibition zone diameters were measured.

Limitations

  • The paper does not include a dedicated limitations section; stated limitations are not given in the paper.
  • Evident from the text: the abstract states an aim to analyze antioxidant ability, but no antioxidant results are reported.
  • Evident from the text: several inconsistencies appear, including 97 isolates versus Table 2 counts summing to 117, CaCO3 listed as 0.5% in the abstract/methods and 1% in the results, molecular identification of only Enterococcus faecalis strain 2358 despite six morphological LAB species, figure captions referring to Pediococcus while the text reports Enterococcus, and antimicrobial table rows labeled “Pl 4.1,” “Pl 4.4,” and “Pl 4.7” without clear explanation.

Model-written summary. Check it against the paper before relying on it.

Integrity signals

Registry

Crossref · 10.31788/rjc.2023.1628239

Cites

19 of 19 references verified0 citation errors · 0 not checkable

Statistics

Nothing to recompute.

Claims the paper makes

0 of 1 cited sources checked · 0 supported · 0 flagged

  • Findingown data

    Isolation of lactic acid bacteria from fresh coconut water using MRSA + CaCO3 selective media yielded 97 isolates.

    “The results showed that 97 isolates were lactic acid bacteria”
    • Also stated in: § RESULTS AND DISCUSSION · plain
      “The result of this isolation, using MRSA + 1% CaCO3 selective media, obtained 97 isolates, as shown in Table -1 below:”
    Quote in the paperSubstantial claim
  • Findingcitation

    According to Bergey's Manual of Systematic Review, the isolates consisted of six species: Lactobacillus plantarum, Lactobacillus brevis, Pediococcus 1, Pediococcus 2, Pediococcus 3, and Pediococcus 4.

    “The Bergey's Manual of Systematic Review showed that the isolates consisted of 6 species such as Lactobacillus plantarum, Lactobacillus brevis, Pediococcus 1, Pediococcus 2, Pediococcus 3, and Pediococcus 4.”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites [9]

      Couldn’t be matched to a paper in Crossref, so it can’t be checked.

  • Findingown data

    More colonies of lactic acid bacteria were obtained at a larger sample dilution.

    “Based on Table-1, more colonies of lactic acid bacteria were obtained at a larger sample dilution.”
    Quote in the paperNot a substantial claim
  • Findingown data

    Molecular identification of the lactic acid bacteria from fresh coconut water showed Enterococcus faecalis strain 2358.

    “Based on Fig. -2, the molecularly identified lactic acid bacteria were Enterococcus faecalis strain 2358.”
    • Also stated in: § Contribution · plain
      “This is because coconut water contains lactic acid bacteria of Enterococcus faecalis strain 2358 and has a good antimicrobial ability.”
    Quote in the paperSubstantial claim
  • Findingown data

    Enterococcus faecalis strain 2358 inhibits S. typhi with the smallest inhibition zone diameter of 23.6 mm.

    “Meanwhile, it can inhibit S. typhi with the smallest inhibition zone diameter of 23.6 mm.”
    Quote in the paperNot a substantial claim
  • May combine two claims. 7
  • Classified as probably not a substantial claim. 1

  • May combine two claims.

    Findingreasoning

    Colonies growing on MRSA media were round, raised and yellowish, while those on MRSA + CaCO3 media grew in the middle of the clear zone.

    “Colonies growing on MRSA media were round, raised, and yellowish, while those on MRSA + Ca CO3 media grew in the middle of the clear zone.”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Findingown data

    Of the isolates, 49 were rod-shaped and the remainder were coccus-shaped.

    “The cell morphology revealed that 49 cells were rod-shaped, and the remainder were coccus-shaped.”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Findingown data

    Morphological, physiological and biochemical tests identified Lactobacillus paracasei, Lactobacillus plantarum, Pediococcus sp 1, Pediococcus sp 2, Pediococcus sp 3, Pediococcus sp 4, and two fungal species, Candida sp and Rizhopus.

    “while morphological, physiological, and biochemical tests showed Lactobacillus paracasei, Lactobacillus plantarum, Pediococcus sp 1, Pediococcus sp 2, Pediococcus sp 3, and Pediococcus sp 4, as well as two species of fungi such as Candida sp and Rizhopus.”
    Quote in the paperSubstantial claim

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Findingown data

    The majority of the colonies were white, with the remainder being cream.

    “The majority of the colonies were white, with the remainder being cream.”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Findingown data

    Lactic acid bacteria isolates from fresh coconut water inhibit the growth of the test bacteria Escherichia coli, Listeria monocytogenes, Salmonella typhi and Staphylococcus aureus, with a large inhibition zone diameter.

    “Based on Table-3, the isolates from fresh coconut water can inhibit the growth of the test bacteria.”
    • Also stated in: § Antimicrobial Activity Analysis · plain
      “The growth of all test bacteria can be inhibited with a large diameter.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    Catalase and motility assays of the isolates yielded negative results.

    “Catalase and motility assays yielded negative results in biochemical studies.”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Findingown data

    Enterococcus faecalis strain 2358 inhibits the growth of L. monocytogenes and E. coli with the largest inhibition zone diameters of 25.3 mm and 25 mm respectively.

    “Enterococcus faecalis strain 2358 can inhibit the growth of L. monocytogenes and E. coli with the largest inhibition zone diameter of 25.3 mm and 25 mm.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Interpretationown data

    Coconut water can be used to treat fever because it contains probiotics and antimicrobial properties, so it is normal for the Minangkabau people of Indonesia to consume it as a supplement drink when they have a fever.

    “Therefore, coconut water can be traditionally used to treat fever medicine because it contains probiotics and antimicrobial properties.”
    • Also stated in: § Contribution · plain
      “Based on the findings, it is normal for the Minangkabau people of Indonesia to consume coconut water as a supplement drink when they have a fever.”
    Quote in the paperSubstantial claim
    • Rests on an assumption: Coconut water has been traditionally used as a fever medicine by the Minangkabau people in Indonesia since ancient times.
8 of 8

  • The paper relies on this without support.

    Coconut water has been traditionally used as a fever medicine by the Minangkabau people in Indonesia since ancient times.

    “Coconut water has been traditionally used as a fever medicine by the Minangkabau people in Indonesia since ancient times, but the chemical or probiotic components have not been widely studied.”

  • The paper relies on this without support.

    Not many studies have isolated lactic acid bacteria in fresh coconut water.

    “Some studies have been conducted on its micro-element content, but not many have isolated lactic acid bacteria in fresh coconut water.”
2 of 2

Self-check

Percentages, sums and reported differences re-computed from the paper’s own numbers.

Each contradiction between two passages listed here was confirmed by an independent check.

  • Table 2 lists 117 isolates, not 97

    Major

    The six isolate counts in Table 2 sum to 117, contradicting the reported total of 97 isolates.

    Reported 97 · recomputed 117

  • CaCO3 concentration differs between methods and results

    Minor

    The abstract gives the MRSA-CaCO3 medium a concentration of 0.5%, while the results describe the isolation medium as containing 1% CaCO3.

  • Species summary substitutes L. brevis for L. paracasei

    Minor

    The text lists Lactobacillus brevis among the morphologically identified species, but Table 2 lists Lactobacillus paracasei instead.

  • White colonies are not the majority in Table 2

    Minor

    Table 2 assigns 49 isolates to white colonies and 68 to cream colonies, contrary to the statement that most colonies were white.

4 of 4

Cites

Flagged first, then by citations

Research works printed but not linked: 1

  1. A. Roy and C. Rai, Bioscience Discovery, 8, 866(2017).

Cited by

Still being checked.

Related

Authors
Suryani, S., Taslim, T., Darmana, I., Rahmawati, R., Arikhman, N., Martinus, B. A., Chrisnawati, C., & Syamsuwirman, S.
Affiliations
  • Universitas Ekasakti (ID)
  • Akademi Farmasi Prayoga Padang (ID)
  • Universitas Bung Hatta (ID)
  • Universitas Baiturrahmah (ID)
  • Universitas Perintis Indonesia (ID)
  • Universitas Mahaputra Muhammad Yamin (ID)
Corresponding author
Suryani, Suryani
ORCID
Venue
RASAYAN Journal of Chemistry
Publisher
Rasayan Journal of Chemistry
Published
2023
Volume
16
Issue
02
Pages
1005-1011
ISSN
  • 0974-1496 (ISSN-L)
  • 0976-0083
DOI
10.31788/rjc.2023.1628239
OpenAlex
W4380050703
Semantic Scholar
35052147218dcd6a425b81d6065dcf151f01e638
Cited by
1
References
19
Citations by year
2024: 1
FWCI
0.13
Type
journal article
Language
en
Open access
Locations
RASAYAN Journal of Chemistry (publishedVersion)
Topics
  • Life Sciences › Agricultural and Biological Sciences › Food Science › Probiotics and Fermented Foods
  • Physical Sciences › Chemistry › Inorganic Chemistry › Coconut Research and Applications
  • Life Sciences › Immunology and Microbiology › Immunology › Aquaculture disease management and microbiota
Keywords
coconut water, Cocos nucifera, lactic acid bacteria, probiotics, Lactobacillus paracasei, Lactobacillus plantarum, Pediococcus, Enterococcus faecalis, antimicrobial activity, antioxidant activity, Staphylococcus aureus, Candida, fever treatment

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