2-Title: Molecular characterization and antimicrobial-resistant pattern of Streptococcus species isolated from bovine mastitis in and around Junagadh district

2-Title: Molecular characterization and antimicrobial-resistant pattern of Streptococcus species isolated from bovine mastitis in and around Junagadh district

Authors: DK Parasana, BB Javia, DT Fefar, DB Barad and SN Ghodasara

Source: Ruminant Science (2021)-10(2):247-252.

How to cite this manuscript: Parasana DK, Javia BB, Fefar DT, Barad DB and Ghodasara SN (2021). Molecular characterization and antimicrobial-resistant pattern of Streptococcus species isolated from bovine mastitis in and around Junagadh district. Ruminant Science 10(2):247-252.


In the present study 98 Streptococcus spp. isolates were recovered from 320 milk samples collected from VCC, KU, Junagadh were used to determine phenotypic and genotypic antimicrobial susceptibility patterns. All Streptococcus spp. isolates were confirmed at the genus level by targeting tuf gene. Str. agalactiae, Str. dysgalactiae and Str. uberis were identified at the species level by targeting 16S rRNA, 16S rRNA and pau gene respectively. Based on PCR detection out of 98 Streptococcus spp. isolates 42, 10 and 16 were Str. agalactiae, Str. dysgalactiae and Str. uberis, respectively. Phenotypic antimicrobial susceptibility was carried out by disc diffusion method and the result showed higher resistance towards ceftriaxone, tetracycline and penicillin and lower resistance towards levofloxacin and gentamicin. For genotypic antimicrobial susceptibility testing ermB, tetK and blaZ genes were targeted. Based on PCR detection ermB, tetK and blaZ genes were found only in 5, 2 and 3 isolates, respectively.


CLSI (2017). Performance standards for antimicrobial susceptibility testing. Clinical and Laboratory Standards Institute (CLSI). NCCLS document M100S 37:19087

Collee JG, Fraser AG, Marmion BP and Simmons DK (2008). In Practical Medical Microbiology. Churchil Livingstone. New York, Edinburgh. London.17th Edn, pp 813-834.

Das Gunjan, Lalnunpuia C, Sarma K, Behera SK, Dutta TK and Bandyopadhyay Samiran (2015). Prevalence of Staphylococcus aureus associated sub-clinical mastitis in crossbred cows in Mizoram. Ruminant Science 4(2):167-170.

Diwakar, Akriti, Choudhary Sunita, Meena Dhirendra, Bhati Taruna and Kataria AK (2019). Antibiotic sensitivity pattern of some Staphylococcus aureus isolates from milk from goats with clinical mastitis. Ruminant Science 8(1):19-22.

Duarte RS, Bruna C, Bellei OP, Miranda1 MA, Brito VP and Teixeira LM (2004). Distribution of serotypes and antimicrobial resistance genes among Streptococcus agalactiae isolates from bovine and human hosts. Journal of Clinical Microbiology 43(12):5899-5906.

FDA (2001). Bacteriological Analytical Manual. Food and Drug Administration (FDA). Cited from http://www.scribd.com, accessed on 08.02.2021

Gao J, Yu FQ, Luo LP, He JZ, Hou RG, Zhang HQ, Li SM, Su JL and Han B (2012). Antibiotic resistance of Streptococcus agalactiae from cows with mastitis. Veterinary Journal 194(3):423-424.

Ghose B and Sharda R (2004). Streptococcal mastitis in dairy cows. Indian Veterinary Medicine Journal 28(2):163-164.

Hegde R (2011). Rapid identification of bacterial pathogens causing subclinical bovine mastitis with special reference to S. aureus, E. coli and predominant Streptococcal species by molecular methods. PhD thesis submitted to KVAFSU, Karnataka.

Khate K and Yadav BR (2010). Incidence of mastitis in Sahiwal cattle and Murrah buffaloes of a closed organized herd. Indian Journal of Animal Science 80:467-469.

Kumar Vaibhav, Patel JS, Patel BR, Mevada VK and Raval AP (2012). Therapeutic efficacy of antimicrobial drugs in clinical mastitis of cross bred cattle. Ruminant Science 1(2):177-180.

Lopardo HA, Vidal P andJeric P (2003). Six-month multicenter study on invasive infections due to group B Streptococci in Argentina. Journal of Clinical Microbiology 41:4688-4694.

Luna VA, Coates P, Eady EA, Cove JH, Nguyen TT and Roberts MC (1999). A variety of gram-positive bacteria carry mobile mef genes. Journal of Antimicrobial Chemotherapy 44(1):19-25.

Miranda PSD, Lannes-Costa PS, Pimentel BAS, Silva LG, Ferreira-Carvalho BT, Menezes GC, Mattos-Guaraldi AL, Hirata JrR, Mota RA and Nagao PE (2018). Biofilm formation on different pH conditions by Streptococcus agalactiae isolated from bovine mastitic milk. Letters in Applied Microbiology 67(3):235-243.

Madushanka DNN, Padmakumara HMS, Kumarasinghe GDN, Sanjeewa MPK and Magamage MPS (2017). Effect of two different bedding systems on udder health management of dairy cows. Ruminant Science 6(1):1-6.

Nithinprabhu K (2010). Isolation, characterization and genetic diversity of Streptococcus species in subclinical bovine mastitis. MVSc Thesis submitted to KVAFSU, Bidar, India.

Picard FJ, Ke D, Boudreau DK, Boissinot M, Huletsky A, Richard D, Ouellette M, Roy PH and Bergeron MG (2004). Use of tuf sequences for genus-specific PCR detection and phylogenetic analysis of 28 streptococcal species. Journal of Clinical Microbiology 42(8):3686-3695.

Pieterse R and Todorov SD (2010). Bacteriocins–exploring alternatives to antibiotics in mastitis treatment. Brazilian Journal of Microbiology 41:542-562.

Raguvaran R, Mondal DB, Jithin MV, Kumar Bipin and Sivakumar M (2016).  Staphylococcus aureus mastitis in a nondescript goat. Ruminant Science 5(2):291-292.

Raorane Abhay, Chothe Shubhadha, Dubal ZB, Barbuddhe SB, Karunakaran M, Doijad Swapnil, Pathak Ajay, Poharkar Krupali and Singh NP (2013). Antimicrobial resistance of the pathogens isolated from bovine mastitis in Goa. Ruminant Science 2(2):139-144.

Rato MG, Bexiga R, Florindo C, Cavaco LM, Vilela CL and Santos-Sanches I (2013). Antimicrobial resistance and molecular epidemiology of streptococci from bovine mastitis. Veterinary Microbiology 161(3-4):286-294.

Riffon R, Sayasith K, Khalil H, Dubreuil P, Drolet M and Lagace J (2001). Development of a rapid and sensitive test for identification of major pathogens in bovine mastitis by PCR. Journal of Clinical Microbiology 39(7):2584-2589.

Savita, Singh AP, Nayak TC, Chahar A, Yadav R and Kachhawa JP (2020). Isolation and identification of bacteria in subclinical mastitis in cattle from Bikaner city. Ruminant Science 9(1):45-48

Shabaz SS, Prameela DR, Sreenivasulu D and Sujatha K (2020). Evaluation of on-farm milk culture system for identification of mastitis pathogens. Ruminant Science 9(2):363-372.

Singh Pawanjit, Nigam Rajesh, Kumar Amit and Pandey Vijay (2018). Isolation and molecular characterization of pathogens associated with mastitis in Sahiwal cows. Ruminant Science 7(1):43-46.

Vesterholm-Nielsen M, Larsen MO, Olsen JE and Aarestrup FM(1999). Occurrence of the blaZ gene in penicillin-resistant Staphylococcus aureus isolated from bovine mastitis in Denmark. Acta Veterinaria Scandinavica 40:279-286.