Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2022 Feb 22:9:784387.
doi: 10.3389/fvets.2022.784387. eCollection 2022.

Whole-Genome Sequence and Interaction Analysis in the Production of Six Enzymes From the Three Bacillus Strains Present in a Commercial Direct-Fed Microbial (Norum™) Using a Bliss Independence Test

Daniel Hernandez-Patlan  1   2 , Bruno Solis-Cruz  1   2 , Juan D Latorre  3 , Ruben Merino-Guzman  4 , Miguel Morales Rodríguez  2 , Catie Ausland  5 , Xochitl Hernandez-Velasco  4 , Oscar Ortiz Holguin  6 , Ramiro Delgado  7 , Billy M Hargis  3 , Pallavi Singh  5 , Guillermo Tellez-Isaias  3
Affiliations

Whole-Genome Sequence and Interaction Analysis in the Production of Six Enzymes From the Three Bacillus Strains Present in a Commercial Direct-Fed Microbial (Norum™) Using a Bliss Independence Test

Daniel Hernandez-Patlan et al. Front Vet Sci. .

Abstract

The three Bacillus strains present in Norum™ were initially selected by their excellent to good relative enzyme activity (REA) production score for amylase, protease, lipase, phytase, cellulase, β-glucanase, and xylanase. Further studies confirmed that the three isolates also showed an antibacterial activity, Gram-positive and Gram-negative poultry pathogens. Norum™ (Eco-Bio/Euxxis Bioscience LLC) is a Bacillus spore direct-fed microbial (DFM). The Bacillus isolates were screened and selected based on in vitro enzyme production profiles. Moreover, in chickens fed high non-starch polysaccharides, this DFM demonstrated to reduce digesta viscosity, bacterial translocation, increase performance, bone mineralization, and balance the intestinal microbiota. In the present study, we present the whole-genome sequence of each of the three isolates in Norum™, as well as the synergistic, additive, or antagonistic effects on the enzyme production behavior of the three Bacillus strains and their combinations when grown together vs. when grown individually. The whole-genome sequence identified isolate AM1002 as Bacillus subtilis (isolate 1), isolate AM0938 as Bacillus amyloliquefaciens (isolate 2), and isolate JD17 as Bacillus licheniformis (isolate 3). The three Bacillus isolates used in the present study produce different enzymes (xylanase, cellulase, phytase, lipase, protease, and β-glucanase). However, this production was modified when two or more Bacillus strains were combined, suggesting possible synergistic, antagonistic, or additive interactions. The Bliss analysis suggested (p < 0.05) that the combination of Bacillus strains 1-2 and 1-2-3 had intermediate effects and predicted that the combination of Bacillus strains 2-3 could have better effects than the combination of all the three Bacillus strains. In summary, the current study demonstrated the need of selecting Bacillus strains based on quantitative enzyme determination and data analysis to assess the impacts of combinations to avoid antagonistic interactions that could limit treatment efficacy. These results suggest that using Bacillus strains 2-3 together could lead to a new generation of DFMs with effects superior to those already examined in Bacillus strains 1-2-3 and, therefore, a potential alternative to growth-promoting antibiotics. More research utilizing poultry models is being considered to confirm and expand the existing findings.

Keywords: Bacillus; bliss analysis; direct-fed microbial; enzymes; genome sequence.

PubMed Disclaimer

Conflict of interest statement

OO is employed by Biodigest S.A.S., and RD is employed by Nutriavícola S.A. The remaining authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.

Figures

Figure 1
Figure 1
Comparison of groups to evaluate the interaction of three strains of Bacillus on the production of xylanase by Bliss independence analysis in single dose with replicates, considering the Tukey test at 95% confidence for comparisons. p < 0.05 indicates significant differences.
Figure 2
Figure 2
Comparison of groups to evaluate the interaction of three strains of Bacillus on the production of cellulase by Bliss independence analysis in single dose with replicates, considering the Tukey test at 95% confidence for comparisons. p < 0.05 indicates significant differences.
Figure 3
Figure 3
Comparison of groups to evaluate the interaction of three strains of Bacillus on the production of phytase by Bliss independence analysis in single dose with replicates, considering the Tukey test at 95% confidence for comparisons. p < 0.05 indicates significant differences.
Figure 4
Figure 4
Comparison of groups to evaluate the interaction of three strains of Bacillus on the production of lipase by Bliss independence analysis in single dose with replicates, considering the Tukey test at 95% confidence for comparisons. p < 0.05 indicates significant differences.
Figure 5
Figure 5
Comparison of groups to evaluate the interaction of three strains of Bacillus on the production of protease by Bliss independence analysis in single dose with replicates, considering the Tukey test at 95% confidence for comparisons. p < 0.05 indicates significant differences.
Figure 6
Figure 6
Comparison of groups to evaluate the interaction of three strains of Bacillus on the production of β-glucanase by Bliss independence analysis in single dose with replicates, considering the Tukey test at 95% confidence for comparisons. p < 0.05 indicates significant differences.
See this image and copyright information in PMC

Similar articles

See all similar articles

Cited by

References

    1. Kurt T, Wong N, Fowler H, Gay C, Lillehoj H, Plummer P, et al. . Strategic priorities for research on antibiotic alternatives in animal agriculture—Results from an expert workshop. Front Vet Sci. (2019) 6:429. 10.3389/fvets.2019.00429 - DOI - PMC - PubMed
    1. Ferdous MF, Arefin MS, Rahman MM, Ripon MMR, Rashid MH, Sultana MR, et al. . Beneficial effects of probiotic and phytobiotic as growth promoter alternative to antibiotic for safe broiler production. J Adv Vet Anim Res. (2019) 6:409–15. 10.5455/javar.2019.f361 - DOI - PMC - PubMed
    1. Hernandez-Patlan D, Solis-Cruz D, Hargis BM, Tellez G. The use of probiotics in poultry production for the control of bacterial infections and aflatoxins. In: Franco-Robles E, Ramírez-Emiliano J, editors. Prebiotics Probiotics-Potential Benefits in Humans Nutrition and Health. London: IntechOpen; (2020). p. 946–1091.
    1. Vreeland RH, Rosenzweig WD, Powers DW. Isolation of a 250 million-year-old halotolerant bacterium from a primary salt crystal. Nature. (2000) 407:897–900. 10.1038/35038060 - DOI - PubMed
    1. Cartman ST, La Ragione RM, Woodward MJ. Bacillus subtilis spores germinate in the chicken gastrointestinal tract. Appl Environ Microbiol. (2008) 74:5254–8. 10.1128/AEM.00580-08 - DOI - PMC - PubMed

LinkOut - more resources