Animal Health and Infectious Diseases

Animal Health and Infectious Diseases

Molecular detection of genes (tetM, tetO, tetL) of Streptococcus agalactiae isolated from milk of goats and sheep of livestock farms around Tehran

Document Type : Original Article

Authors
Sepideh Assadi 1
Shirin MohammadiPour 2
Erfan ZohourianPordel 3
MohammadHosein Ghaffari 3
Farzad Esfandiyar 4
Bahar NayeriFasaei 5
1 Ph.D. in Bacteriology, Faculty of Veterinary Medicine, University of Tehran, Tehran, Iran
2 Ph.D. student of Bacteriology, Faculty of Veterinary Medicine, Bahnar University, Kerman, Iran
3 DVM, student of Veterinary Medicine, Research Sciences Unit, Islamic Azad University, Tehran, Iran
4 DVM, student of veterinary medicine, Faculty of Veterinary Medicine, Bahnar University, Kerman, Iran
5 Associate Professor, Department of Microbiology and Immunology, Faculty of Veterinary Medicine, University of Tehran, Tehran, Iran
10.22034/jahid.2024.710959
Abstract
Background and Aim: The condition of inflammation of the mammary tissue, often referred to as mastitis which can
occur due to various factors, with Streptococcus agalactiae being one of the significant microbial agents responsible
for its development. It often causes treatment costs and premature culling. The present study aimed to investigate
antibiotic resistance genes for oxytetracycline in Streptococcus agalactiae strains isolated from the milk of goats and
sheep in the Tehran region.
Materials and Methods: A total of 240 milk samples from goats and sheep suffering from mastitis were collected
from industrial farms around Tehran and cultured on blood agar medium. The grown colonies underwent standard
phenotypic and biochemical tests. Subsequently, the antibiotic sensitivity of the isolates was studied using the KirbyBauer disk diffusion method. To confirm the identification of bacteria from pure cultures confirmed by biochemical
tests, PCR was employed for Streptococcus agalactiae. Finally, the resistance genes (tetO, tetM, tetL) were examined.
Results: The results indicated that out of the total 240 milk samples, 8 samples (6.7%) and 14 samples (11.7%) were
infected with Streptococcus agalactiae in goats and sheep, respectively. The investigation of antibiotic resistance genes
revealed that all positive samples from sheep's milk contained resistance genes to oxytetracycline. Among the 8
positive samples in goat's milk, only 4 samples were found to contain the tetM gene.
Conclusion: Considering the presence of a high number of antibiotic resistance genes, proper use of antibiotics and
continuous and rapid screening of this microorganism should receive more attention to prevent the emergence and
spread of antibiotic-resistant Streptococcus agalactiae strains

Keywords
Streptococcus agalactiae
mastitis
cow
tetracycline
Subjects
  1. Levy, S.B., The challenge of antibiotic resistance. Scientific American, 1998. 278(3): p. 46-53. 2. P
  2. oyart, C., et al., Genetic Basis of Antibiotic Resistance in Streptococcusagalactiae Strains Isolated in a French Hospital. Antimicrobial Agents and Chemotherapy, 2003. 47(2): p. 794-797. 
  3. Furfaro, L.L., B.J. Chang, and M.S. Payne, Perinatal Streptococcus agalactiae epidemiology and surveillance targets. Clinical microbiology reviews, 2018. 31(4): p. 10.1128/cmr. 00049-18.
  4. Seale, A.C., et al., Maternal colonization with Streptococcus agalactiae and associated stillbirth and neonatal disease in coastal Kenya. Nature microbiology, 2016. 1(7): p. 1-10. 
  5. Sambola, A., et al., Streptococcus agalactiae Infective Endocarditis: Analysis of 30 Cases and Review of the Literature, 1962–1998. Clinical Infectious Diseases, 2002. 34(12): p. 1576-1584. 
  6. Basdew, I. and M. Laing, Mini-Review: Biological control of bovine mastitis using bacteriophage therapy. Science against microbial pathogens: communicating current research and technological advances, 2011: p. 386-393. 
  7. Richards, V.P., et al., Comparative genomics and the role of lateral gene transfer in the evolution of bovine adapted Streptococcus agalactiae. Infection, Genetics and Evolution, 2011 :)6(11 .p. 1263-1275. 
  8. Radtke, A., et al., Multiple-locus variant-repeat assay (MLVA) is a useful tool for molecular epidemiologic analysis of Streptococcus agalactiae strains causing bovine mastitis. Veterinary microbiology, 2012. 157(3- 4): p. 398-404.
  9. Breeding, K.M., et al., Real-time PCR-based serotyping of Streptococcus agalactiae. Scientific reports, 2016. 6(1): p. 38523. 
  10. Imperi, M., et al., A multiplex PCR assay for the direct identification of the capsular type (Ia to IX) of Streptococcusagalactiae. Journal of microbiological methods, 2010. 80(2): p. 212-214.
  11. Rosini, R. and I. Margarit, Biofilm formation by Streptococcus agalactiae: influence of environmental conditions and implicated virulence factors. Frontiers in cellular and infection microbiology, 2015. 5: p. 6.
  12. Ebrahimi, A., et al. Biofilm formation, hemolysin production and antimicrobial susceptibilities of Streptococcus agalactiae isolated from the mastitis milk of dairy cows in Shahrekord district, Iran. in Veterinary Research Forum: an International Quarterly Journal. 2013. Faculty of Veterinary Medicine, Urmia University, Urmia, Iran.
  13. Miranda, P., et al., Biofilm formation on different pH conditions by Streptococcus agalactiae isolated from bovine mastitic milk. Letters in Applied Microbiology, 2018. 67(3): p. 235-243.
  14. Moatamedih, et al., A polymerase chain reaction based study on the subclinical mastitis caused by Streptococcus agalactiae, S. dysgalactiae and S. uberis in cattle in Ahvaz. Iranian Journal of Veterinary Research, 2007. 8(3):260-265.Ůž
  15. Reza R., et al., Antibacterial effect of essential oils of peppermint and pennyroyal on major bovine mastitis
    bacteria. New Findings in Veterinary Microbiology, 2023; 1(6): 64-75.
  16. Pinto, T.C.A., et al., Conjugative transfer of resistance determinants among human and bovine Streptococcus agalactiae. Brazilian Journal of .Microbiology, 2014. 45: p. 785-789
  17. Dogan, B., et al., Distribution of serotypes and antimicrobial resistance genes among Streptococcus agalactiae isolates from bovine and human hosts. Journal of clinical microbiology, 2005. 43(12): p. 5899- 5906.
  18. Soltani M., et al., Antibiotic Resistance in Pathogenic Bacteria the Causative Agents of Bacterial Diseases in Farmed Rainbow Trout (Onchorhynchus mykiss) in Iran. Journal of Veterinary Research, 2023; 2 (78): 5-9.
  19. Asadi, S., et al., Antibacterial and anti-biofilm properties of carvacrol alone and in combination with cefixime against Escherichia coli. BMC Microbiology, 2023. 23(1): p. 55.
  20. Meiri-Bendek ,I., et al., A PCR-Based Method for the Detectionof Streptococcus agalactiae in Milk. Journal of Dairy Science, 2002. 85(7): p. 1717-1723.
  21. Duarte, R.S., et al., Phenotypic and molecular characteristics of Streptococcus agalactiae isolates recovered from milk of dairy cows in Brazil. J Clin Microbiol, 2004. 42(9): p. 4214-22.
  22.  Silva, J., et al., In vitro antimicrobial susceptibility and genetic resistance determinants of Streptococcus agalactiae isolated from mastitic cows in Brazilian dairy herds.Semina: Ciências Agrárias, 2017. 38: p. 2581.
  23. Momtaz, H., et al., Molecular detection of Streptococcus uberis and Streptococcus agalactiae in the mastitic cows milks in Isfahan province. Biological Journal of Microorganism, 2012. 1(2): p. 71-76.
  24. Brown, M. and M. Roberts, Tetracycline resistance determinants in streptococcal species isolated from the bovine mammary gland. Veterinary microbiology, 1991. 29(2): p. 173-180.
  25. Schwarz, S., I. Wibawan, and C. Lämmler, Distribution of genes conferringcombined resistance to tetracycline and minocycline among group B streptococcal isolates from humans and various animals. Zentralblatt für Bakteriologie, 1994. 281(4): p. 526-533.
  26. Guérin-Faublée, V., et al., Antimicrobial susceptibility of Streptococcus species isolated from clinical mastitis in dairy cows. International journal of antimicrobial agents, 2002. 19(3): p. 219-226. 
  27. Rato, M.G., et al., Antimicrobial resistance and molecular epidemiology of streptococci from bovine mastitis. Veterinarymicrobiology, 2013. 161(3-4): p. 286-294.
  28. Lindeman, C.J., et al., Susceptibility to antimicrobial agents among bovine mastitis pathogens isolated from North American dairy cattle, 2002–2010. Journal of Veterinary Diagnostic Investigation, 2013. 25 :)5(p. 581-591. 
  29. Bolukaoto, J.Y., et al., Antibiotic resistance of Streptococcus agalactiae isolated from pregnant women in Garankuwa, South Africa. BMC research notes, 2015. 8(1): p. 1-7. 
  30. Gao, J., et al., Antibiotic resistance of Streptococcus agalactiae from cows with mastitis. The Veterinary Journal, 2012. 194(3): p. 423-424.
  31. Duarte, R.S., et al., Phenotypic and molecular characteristics of Streptococcus agalactiae isolates recovered from milk of dairy cows in Brazil. Journal of clinical microbiology, 2004. 42(9): p. 4214-4222.
  32. Duarte, R.S., et al., Distribution of antimicrobial resistance and virulence-related genes among Brazilian group B streptococci recovered from bovine and human sources. Antimicrobial Agents and Chemotherapy, 2005. 49 :)1(p. 97-103.
  33. Dutra, V.G., et al., Streptococcus agalactiae in Brazil: serotype distribution, virulence determinants and antimicrobial susceptibility. BMC infectious diseases, 2014. 14: p. 1-9.
  34. Chopra, I. and M. Roberts, Tetracycline antibiotics : mode of action, applications, molecular biology, and epidemiology of bacterial resistance. Microbiology and molecular biology reviews, 2001. 65(2): p. 232-260. 
  35. Karami P. et al., Determination Pattern of Antibiotic Resistance in Entropathogenic Escherichia coli Strains Isolated from Children with Diarrhea. Avicenna Journal Clinical Medicine, 2012; 19(1): 27-31.
Animal Health and Infectious Diseases
Volume 1, Issue 1 - Serial Number 1
December 2024
Pages 17-25