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. 2023 Jan 19;18(1):e0280412.
doi: 10.1371/journal.pone.0280412. eCollection 2023.

Identification and characterization of probiotics isolated from indigenous chicken (Gallus domesticus) of Nepal

Mohan Gupta  1 , Roji Raut  2 , Sulochana Manandhar  2 , Ashok Chaudhary  2 , Ujwal Shrestha  1 , Saubhagya Dangol  1 , Sudarshan G C  1   3 , Keshab Raj Budha  4 , Gaurab Karki  5   6 , Sandra Díaz-Sánchez  7 , Christian Gortazar  7 , José de la Fuente  7   8 , Pragun Rajbhandari  2 , Prajwol Manandhar  2 , Rajindra Napit  2   9 , Dibesh Karmacharya  2   9   10
Affiliations

Identification and characterization of probiotics isolated from indigenous chicken (Gallus domesticus) of Nepal

Mohan Gupta et al. PLoS One. .

Abstract

Background: Excessive and irrational use of antibiotics as growth promoters in poultry has been one of key factors contributing to increased emergence of antibiotics resistant bacteria. Several alternatives for antibiotic growth promoters are being sought, and the search for effective probiotics to be used as feed additives is amongst the promising ones. Our study aimed to isolate and test potential probiotics bacteria from cloacal swabs of various indigenous chicken (Gallus domesticus) breeds from rural outskirts of the Kathmandu valley (Nepal).

Methods: Selective isolation of probiotics was conducted by micro-aerophilic enrichment of sample in MRS Broth at 37°C, followed by culturing on MRS agar supplemented with 5 g/L of CaCO3. Isolated bacterial colonies producing transparent halo were selected as potential lactic acid bacteria (LAB), and tested for their antibacterial activity, phenotypic and biochemical characteristics, acidic yield, and tolerance to acid and bile.

Results: A total of 90 potential LAB were isolated from cloacal samples collected from 41 free-ranging chickens of indigenous breeds. Of these, 52 LAB isolates (57%) showed variable antibacterial activity to at least one bacterial pathogen. Of 52 LAB, 46 isolates fulfilled phenotypic and biochemical criteria of Lactobacillus spp. Of these, 37 isolates produced varying percentage yields of lactic acid, 27 isolates showed survival at pH 3.0, and 17 isolates showed survival tolerances in the presence of 0.3% and 0.5% bile salts for 24 hours. Phylogenetic analysis of 16S rDNA sequencing of LAB isolates fulfilling in vitro probiotics properties showed that 3 isolates had genetic identity of 99.38% with Lactobacillus plantarum, while one isolate was genetically similar (99.85%) with the clade of L. reuteri, L. antri and L. panis.

Conclusion: Our study identified four Lactobacillus spp. strains having potential probiotics properties. Further investigations are needed to evaluate these isolates to be used as poultry probiotics feed supplement.

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Conflict of interest statement

The authors have declared that no competing interests exist.

Figures

Fig 1
Fig 1. The map of sampling districts (yellow colored) of Nepal showing the sampling locations indicated as numbered circles.
The sample locations are as follows: 1) Lakuribhanjyang, 2) Parthali- Bethankchowk, 3) Chalnakhel-Bosandanda and 4) Jhor- Dhakalchaur. The diameter of sampling locations are shown as varying proportionately with the number of samples collected from that location as indicated in the figure legend.
Fig 2
Fig 2. Maximum likelihood tree based on 16S rDNA gene sequences of different Lactobacillus spp.
The isolates of the study are indicated in red font. Lactococcus plantarum was chosen as an out-group. The scale bar (0.03) shows the nucleotide substitution rate per site. Bootstrap probabilities as determined for 1000 replicates are shown at nodes. The isolate C4/36(4) has been labeled as C4 in the given phylogenetic tree.
See this image and copyright information in PMC

References

    1. FAO. Probiotics in animal nutrition: production, impact and regulation. Bajagai YS, Klieve AV, Dart PJ, Bryden WL, editors. Rome: FAO Animal Production and Health; 2016.
    1. Van TTH, Yidana Z, Smooker PM, Coloe PJ. Antibiotic use in food animals worldwide, with a focus on Africa: Pluses and minuses. Journal of global antimicrobial resistance. 2020;20:170–7. doi: 10.1016/j.jgar.2019.07.031 - DOI - PubMed
    1. Poudel U, Dahal U, Upadhyaya N, Chaudhari S, Dhakal S. Livestock and poultry production in Nepal and current status of vaccine development. Vaccines. 2020;8(2):322. doi: 10.3390/vaccines8020322 - DOI - PMC - PubMed
    1. Jensen HA, editor. Paradigm and visions: network for poultry production and health in developing countries. Proceedings of a Workshop on Poultry as a tool in poverty eradication and promotion of gender equality, Tune Landboskole, Denmark; 1999.
    1. Hosain MZ, Kabir SL, Kamal MM. Antimicrobial uses for livestock production in developing countries. Veterinary World. 2021;14(1):210. doi: 10.14202/vetworld.2021.210-221 - DOI - PMC - PubMed

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