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Phenotypic and genotypic of gram-positive coccus β-hemolysis bacteria isolates from tonsil and nasal of pig and antimicrobial susceptibility test against penicillin G and tetracycline

  • Komang Januartha Putra Pinatih ,
  • Komang Tri Astuti ,
  • I Wayan Suardana ,
  • I Made Sukada ,
  • Siti Isrina Oktavia Salasia ,
  • I Gede Putu Supadmanaba ,
  • Desak Made Wihandani ,


Background: The nasal cavity of a pig serves as an entry point and a habitat for the colonization of commensal microbes and pathogenic bacteria such as bacterial zoonosis. This study aimed to identify the phenotypic and genotypic of Gram-positive β-hemolytic organisms collected from nasal and tonsil swabs of pigs that were collected from the abattoir surrounding outbreak area, complete with testing sensitivity test of the bacterial isolates to Penicillin G and Tetracycline antibiotics.

Material and Methods:  Totaling 18 isolates consisting of 9 isolates from nasal and 9 isolates from tonsil of coccus β-Hemolysis bacteria originating from 60 samples were used. These isolates were then conventionally identified, then molecularly using 16S rRNA gene analysis. The sensitivity test was carried out by the Kirby Bauer method following the standards of the Clinical and Laboratory Standards Institute (CLSI).

Results: The results of the phenotypic identification (biochemical test) revealed that 9 out of 18 isolates were identified as Enterococcus spp. Furthermore, the confirmation of isolates by molecular analysis i.e. the 16S rRNA gene showed consistency with the biochemical test,  and there was confirmed as Enterococcus faecium and Enterococcus faecalis. The sensitivity test showed as many as 9 isolates resistant to Penicillin G and Tetracycline,  5 isolates were resistant, 1 isolate was sensitive and 3 were intermediates. Moreover, the results of the sensitivity test to Penicillin G and Tetracycline, 5 out of 9 isolates showed resistance to more than one antibiotic.

Conclusion: The phenotypic and genotypic of Gram-Positive Coccus β-Hemolysis bacteria isolates from the tonsil and nasal of pigs showed as Enterococcus faecium and Enterococcus faecalis. The use of antibiotics such as Penicillin G and Tetracycline is not recommended for the treatment of this agent resulting in most of the agents being resistant.


  1. Eddicks M, Eddicks L, Stadler J, Hermanns W, Ritzmann M. The porcine respiratory disease complex (PRDC) - a clinical review. Tierarztl Prax Ausg G Grosstiere Nutztiere. 2021;49(2):120–32.
  2. Naomi C, Suardana IW, Suarsana IN. Isolated Hemolysis Profile of Streptococcus Sp. Isolation Result from Swine’s Tonsil In Slaughter House at Punggul and Bongkasa Village. Journal of Veterinary and Animal Sciences. 2019;2(2):46. Available from:
  3. Yanti NLMS, Suardana IW, Suarjana IGK, Suarjana IGK. Hemolytic Profile Of Streptococcus Spfrom Nasal Swab Isolation At Traditional Farm In Bongkasa Village, Abiansemal Subdistrict, Badung Regency, Bali. Journal of Veterinary and Animal Sciences. 2019;2(2):52.
  4. Suharsono H, Suardana IW, Putri RK. Identification of PST 10 bacterial isolate with β-hemolysis characteristic isolated from pig’s tonsil. Bali Medical Journal. 2022;11(1):56–60.
  5. Moges F, Endris M, Belyhun Y, Worku W. Isolation and characterization of multiple drug resistance bacterial pathogens from wastewater in hospital and non-hospital environments, Northwest Ethiopia. BMC Res Notes. 2014;7(1).
  6. Wardoyo EH, Suardana IW, Yasa IWPS, Sukrama IDM. Antibiotics susceptibility of Escherichia coli isolates from clinical specimens before and during the COVID-19 pandemic. Iran J Microbiol. 2021;13(2):156–60. Available from:
  7. Suardana IW. Analysis of Nucleotide Sequences of the 16S rRNA Gene of Novel Escherichia coli Strains Isolated from Feces of Human and Bali Cattle. J Nucleic Acids. 2014/09/09. 2014;2014:475754. Available from:
  8. Sukrama IDM, Praja RK, Fatmawati NND. Pheno-genotypic profile of Vibrio choleraehemolysin (hlyA) isolated from shrimp and shellfish at the Kedonganan fish market, Bali-Indonesia. Bali Medical Journal. 2017;5(2):366–9. Available from:
  9. Pinatih KJP, Suardana IW, Sukrama IDM, Swacita IBN, Putri RK. Biochemical and molecular identification of Gram-positive isolates with β-hemolysis activity isolated from the nasal swab of pigs during the human meningitis outbreak in Badung Regency, Bali-Indonesia. Vet World. 2022/01/25. 2022;15(1):140–6. Available from:
  10. Suardana IW, Dinarini NMAA, Sukrama IDM. Identifikasi Spesies Streptokokus ?-Hemolisis Hasil Isolasi dari Nasal dan Tonsil Babi dengan Uji Basitrasin. Buletin Veteriner Udayana. 2021;27. Available from:
  11. Suardana IW. Erratum to “Analysis of Nucleotide Sequences of the 16S rRNA Gene of Novel Escherichia coli Strains Isolated from Feces of Human and Bali Cattle.” J Nucleic Acids. 2014/12/29. 2014;2014:412942. Available from:
  12. Suardana IW, Pinatih KJP, Widiasih DA, Artama WT, Asmara W, Daryono BS. Regulatory elements of stx2 gene and the expression level of Shiga-like toxin 2 in Escherichia coli O157:H7. Journal of Microbiology, Immunology and Infection. 2018;51(1):132–40. Available from:
  13. Tamura K, Stecher G, Kumar S. MEGA11: Molecular Evolutionary Genetics Analysis Version 11. Mol Biol Evol. 2021;38(7):3022–7. Available from:
  14. Janda JM, Abbott SL. 16S rRNA gene sequencing for bacterial identification in the diagnostic laboratory: pluses, perils, and pitfalls. J Clin Microbiol. 2007/07/11. 2007;45(9):2761–4. Available from:
  15. Liu Z, Zheng H, Gottschalk M, Bai X, Lan R, Ji S, et al. Development of multiplex PCR assays for the identification of the 33 serotypes of Streptococcus suis. PLoS One. 2013;8(8):e72070–e72070. Available from:
  16. CLSI. M100 Performance Standards for Antimicrobiafile:///C:/Users/K/Downloads/Documents/2015_art_esprmartins1.pdfl. 2021.
  17. Semedo T, Almeida Santos M, Martins P, Silva Lopes MF, Figueiredo Marques JJ, Tenreiro R, et al. Comparative study using type strains and clinical and food isolates to examine hemolytic activity and occurrence of the cyl operon in enterococci. J Clin Microbiol. 2003;41(6):2569–76. Available from:
  18. Adams L, Boopathy R. Isolation and characterization of enteric bacteria from the hindgut of Formosan termite. Bioresour Technol. 2005;96(14):1592–8. Available from:
  19. Koeth LM, DiFranco JM. Comparison of daptomycin Etest MICs on Mueller Hinton, IsoSensitest and brain heart infusion agars from Europe against 20 Staphylococcus aureus isolates. European Journal of Clinical Microbiology & Infectious Diseases. 2010;29(10):1261–4. Available from:
  20. Stasiewicz MJ, Wiedmann M, Bergholz TM. The Combination of Lactate and Diacetate Synergistically Reduces Cold Growth in Brain Heart Infusion Broth across Listeria monocytogenes Lineages. J Food Prot. 2010;73(4):631–40. Available from:
  21. Shenoy S, Mala K. Enterococcus Faecalis: An Endodontic Pathogen [Internet]. 2006. Available from:
  22. Martins Teixeira L, Da Glória M, Carvalho S, Facklam RR, Shewmaker PL. Manual of Clinical Microbiology Enterococcus. In: Manual of Clinical Microbiology, 11th Edition. 2015. Available from:
  23. Sacchi CT, Whitney AM, Mayer LW, Morey R, Steigerwalt A, Boras A, et al. Sequencing of 16S rRNA gene: a rapid tool for identification of Bacillus anthracis. Emerg Infect Dis. 2002;8(10):1117–23. Available from:
  24. Klein G. Taxonomy, ecology and antibiotic resistance of enterococci from food and the gastro-intestinal tract. Int J Food Microbiol. 2003;88(2–3):123–31. Available from:
  25. Mohanty S, Singhal R, Sood S, Dhawan B, Kapil A, Das BK. Citrobacter infections in a tertiary care hospital in Northern India. Journal of Infection. 2007;54(1):58–64. Available from:
  26. Rosa R, Creti R, Venditti M, D’Amelio R, Arciola CR, Montanaro L, et al. Relationship between biofilm formation, the enterococcal surface protein (Esp) and gelatinase in clinical isolates ofEnterococcus faecalisandEnterococcus faecium. FEMS Microbiol Lett. 2006;256(1):145–50. Available from:
  27. Munita JM, Arias CA. Mechanisms of Antibiotic Resistance. Microbiol Spectr. 2016;4(2):10.1128/microbiolspec.VMBF-0016–2015. Available from:
  28. Signoretto C, Lleò MM, Tafi MC, Canepari P. Cell wall chemical composition of Enterococcus faecalis in the viable but nonculturable state. Appl Environ Microbiol. 2000;66(5):1953–9. Available from:
  29. Allen HK, Stanton TB. Altered Egos: Antibiotic Effects on Food Animal Microbiomes. Annu Rev Microbiol. 2014;68(1):297–315. Available from:
  30. Velhner M, Milanov D. Resistance to tetracycline in Escherichia coli and Staphylococcus aureus: brief overview on mechanisms of resistance and epidemiology. Archives of Veterinary Medicine. 2016;8(1):27–36. Available from:
  31. Ogawara H. Comparison of Antibiotic Resistance Mechanisms in Antibiotic-Producing and Pathogenic Bacteria. Molecules. 2019;24(19):3430. Available from:
  32. Pratiwi RH. Mekanisme pertahanan bakteri patogen terhadap antibiotik. Jurnal Pro-Life. 2017;4:418–29.
  33. van der Meer JWM, Gyssens IC. Quality of antimicrobial drug prescription in hospital. Clinical Microbiology and Infection. 2001;7(s6):12–5. Available from:
  34. Adebowale OO, Adeyemo OK, Awoyomi O, Dada R, Adebowale O. Antibiotic use and practices in commercial poultry laying hens in Ogun State Nigeria. Rev Elev Med Vet Pays Trop. 2016;69(1):41–5. Available from:
  35. Bester LA, Essack SY. Observational Study of the Prevalence and Antibiotic Resistance of Campylobacter spp. from Different Poultry Production Systems in KwaZulu-Natal, South Africa. J Food Prot. 2012;75(1):154–9. Available from:
  36. Van Boeckel TP, Brower C, Gilbert M, Grenfell BT, Levin SA, Robinson TP, et al. Global trends in antimicrobial use in food animals. Proc Natl Acad Sci U S A. 2015/03/19. 2015;112(18):5649–54. Available from:
  37. Natadidjaja RI, Kusuma AS, Sudradjad GB, Nugrohowati L. The Association between Medical History-based Risks and Sepsis Events in Immunocompromised Patients according to Type III Stratification of the Indonesian Regulation on the Prospective Antimicrobial System (Regulasi Antimikroba Sistem Prospektif / RASPRO). Bali Medical Journal. 2021;10(3):1031–6.
  38. Gyles C, Boerlin P. Horizontally Transferred Genetic Elements and Their Role in Pathogenesis of Bacterial Disease. Vol. 51, Veterinary Pathology. 2014. p. 328–40.

How to Cite

Pinatih, K. J. P., Astuti, K. T. ., Suardana, I. W. ., Sukada, I. M. ., Salasia, S. I. O. ., Supadmanaba, I. G. P. ., & Wihandani, D. M. . (2023). Phenotypic and genotypic of gram-positive coccus β-hemolysis bacteria isolates from tonsil and nasal of pig and antimicrobial susceptibility test against penicillin G and tetracycline. Bali Medical Journal, 12(3), 2407–2414.




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Komang Januartha Putra Pinatih
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Komang Tri Astuti
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I Wayan Suardana
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I Made Sukada
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Siti Isrina Oktavia Salasia
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I Gede Putu Supadmanaba
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Desak Made Wihandani
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