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Journal of Integrative Agriculture  2020, Vol. 19 Issue (11): 2784-2791    DOI: 10.1016/S2095-3119(20)63185-9
Special Issue: 动物医学合辑Veterninary Medicine
Animal Science · Veterinary Medicine Advanced Online Publication | Current Issue | Archive | Adv Search |
Detection of antimicrobial resistance and virulence-related genes in Streptococcus uberis and Streptococcus parauberis isolated from clinical bovine mastitis cases in northwestern China
ZHANG Hang, YANG Feng, LI Xin-pu, LUO Jin-yin, WANG Ling, ZHOU Yu-long, YAN Yong, WANG Xu-rong, LI Hong-sheng
Lanzhou Institute of Husbandry and Pharmaceutical Sciences, Chinese Academy of Agricultural Sciences/Key Lab of Veterinary Pharmaceutical Development, Ministry of Agriculture and Rural Affairs/Engineering and Technology Research Center of Traditional Chinese Veterinary Medicine, Lanzhou 730050, P.R.China
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The objectives of this study were to investigate antimicrobial resistance of Streptococcus uberis and Streptococcus parauberis isolated from cows with bovine clinical mastitis in China and to examine the distribution of resistance- and virulence-related gene patterns.  Antimicrobial susceptibility was determined by the E-test.  Genes encoding antimicrobial resistance and invasiveness factors were examined by PCR.  A total of 27 strains were obtained from 326 mastitis milk samples.  Streptococcus parauberis isolates (n=11) showed high resistance to erythromycin (90.9%), followed by tetracycline (45.5%), chloramphenicol (36.4%) and clindamycin (27.3%).  Streptococcus uberis isolates (n=16) were highly resistant to tetracycline (81.3%) and clindamycin (62.5%).  Both species were susceptible to ampicillin.  The most prevalent resistance gene in S. uberis was tetM (80.0%), followed by blaZ (62.5%) and ermB (62.5%).  However, tetM, blaZ, and ermB genes were only found in 27.3, 45.5, and 27.3%, respectively, of S. parauberis.  In addition, all of the isolates carried at least one selected virulence-related gene.  The most prevalent virulence-associated gene pattern in the current study was sua+pauA/skc+gapC+hasC detected in 22.2% of the strains.  One S. uberis strain carried 7 virulence-associated genes and belonged to the sua+pauA/skc+gapC+cfu+hasA+hasB+hasC pattern.  More than 59.3% of analysed strains carried 4 to 7 virulence-related genes.  Our findings demonstrated that S. parauberis and S. uberis isolated from clinical bovine mastitis cases in China exhibited diverse molecular ecology, and that the strains were highly resistant to antibiotics commonly used in the dairy cow industry.  The data obtained in the current study contribute to a better understanding of the pathogenesis of bacteria in mastitis caused by these pathogens, and the findings are relevant to the development of multivalent vaccines and targeted prevention procedures.
Received: 11 October 2019   Accepted:
Fund: This study was supported by the National Key Research and Development Project of China (2017YFD0502200), the National Natural Science Foundation of China (31802232), and the International Science and Technology Cooperation Project of Gansu Province, China (17YF1WA169).
Corresponding Authors:  Correspondence WANG Xu-rong, Tel: +86-931-2112161, Fax: +86-931-2114180, E-mail:; LI Hong-sheng, Tel: +86-931-2164183, Fax: +86-931-2114180, E-mail:   
About author:  ZHANG Hang, E-mail:, Mobile: +86-16602725014;

Cite this article: 

ZHANG Hang, YANG Feng, LI Xin-pu, LUO Jin-yin, WANG Ling, ZHOU Yu-long, YAN Yong, WANG Xu-rong, LI Hong-sheng. 2020. Detection of antimicrobial resistance and virulence-related genes in Streptococcus uberis and Streptococcus parauberis isolated from clinical bovine mastitis cases in northwestern China. Journal of Integrative Agriculture, 19(11): 2784-2791.

Almeida R A, Dego O K, Headrick S I, Lewis M J, Oliver S P. 2015. Role of Streptococcus uberis adhesion molecule in the pathogenesis of Streptococcus uberis mastitis. Veterinary Microbiology, 179, 332–335.
Boireau C, Cazeau G, Jarrige N, Calavas D, Madec J Y, Leblond A, Haenni M, Gay E. 2018. Antimicrobial resistance in bacteria isolated from mastitis in dairy cattle in France, 2006–2016. Journal of Dairy Science, 101, 9451–9462.
Cameron M, Saab M, Heider L, Mcclure J T, Rodriguez-Lecompte J C, Sanchez J. 2016. Antimicrobial susceptibility patterns of environmental Streptococci recovered from bovine milk samples in the Maritime Provinces of Canada. Frontiers in Veterinary Science, 3, 79.
Chuzeville S, Puymege A, Madec J Y, Haenni M, Payot S. 2012. Characterization of a new CAMP factor carried by an integrative and conjugative element in Streptococcus agalactiae and spreading in streptococci. PLoS ONE, 7, e48918.
CLSI (Clinical and Laboratory Standards Institute). 2013. Performance Standards for Antimicrobial Susceptibility Testing: Twenty-Third Informational Supplement. M100-S23. CLSI, Wayne, PA.
Fernandez-No I C, Bohme K, Calo-Mata P, Canas B, Gallardo J M, Barros-Velazquez J. 2012. Isolation and characterization of Streptococcus parauberis from vacuum-packaging refrigerated seafood products. Food Microbiology, 30, 91–97.
Field T R, Ward P N, Pedersen L H, Leigh J A. 2003. The hyaluronic acid capsule of Streptococcus uberis is not required for the development of infection and clinical mastitis. Infection and immunity, 71, 132–139.
František Z, Vasi? M, Ele?ko J, Zigová M, Farkašová Z. 2017. Mastitis pathogens isolated from samples of milk in dairy cows and their resistance against antimicrobial agents. Journal of Food Science and Engineering, 7, 110–113.
Han S Y, Kang B K, Kang B J, Shin S P, Soen B H, Kim J M, Kim J H, Choresca Jr C H, Han J E, Jun J W, Park S C. 2011. Prevalence and different characteristics of two serotypes of Streptococcus parauberis isolated from the farmed olive flounder, Paralichthys olivaceus (Temminck and Schlegel), in Korea. Journal of Fish Diseases, 34, 731–739.
Ip M, Lyon D J, Yung R W, Chan C, Cheng A F. 2001. Macrolide resistance in Streptococcus pneumoniae in Hong Kong. Antimicrobial Agents and Chemotherapy, 45, 1578–1580.
De Jong A, Garch F E, Simjee S, Moyaert H, Rose M, Youala M, Siegwart E, Vetpath Study G. 2018. Monitoring of antimicrobial susceptibility of udder pathogens recovered from cases of clinical mastitis in dairy cows across Europe: VetPath results. Veterinary Microbiology, 213, 73–81.
Kaczorek E, Malaczewska J, Wojcik R, Rekawek W, Siwicki A K. 2017a. Phenotypic and genotypic antimicrobial susceptibility pattern of Streptococcus spp. isolated from cases of clinical mastitis in dairy cattle in Poland. Journal of Dairy Science, 100, 6442–6453.
Kaczorek E, Malaczewska J, Wojcik R, Siwicki A K. 2017b. Biofilm production and other virulence factors in Streptococcus spp. isolated from clinical cases of bovine mastitis in Poland. BMC Veterinary Research, 13, 398.
Khan I U, Hassan A A, Abdulmawjood A, Lammler C, Wolter W, Zschock M. 2003. Identification and epidemiological characterization of Streptococcus uberis isolated from bovine mastitis using conventional and molecular methods. Journal of Veterinary Science, 4, 213–224.
Kuipers A, Koops W J, Wemmenhove H. 2016. Antibiotic use in dairy herds in the Netherlands from 2005 to 2012. Journal of Dairy Science, 99, 1632–1648.
Luther D A, Almeida R A, Oliver S P. 2008. Elucidation of the DNA sequence of Streptococcus uberis adhesion molecule gene (sua) and detection of sua in strains of Streptococcus uberis isolated from geographically diverse locations. Veterinary Microbiology, 128, 304–312.
Malhotra-Kumar S, Lammens C, Piessens J, Goossens H. 2005. Multiplex PCR for simultaneous detection of macrolide and tetracycline resistance determinants in streptococci. Antimicrobial Agents and Chemotherapy, 49, 4798–4800.
McDonald W L, Fry B N, Deighton M A. 2005. Identification of Streptococcus spp. causing bovine mastitis by PCR-RFLP of 16S–23S ribosomal DNA. Veterinary Microbiology, 111, 241–246.
Van Der Merwe J, Prysliak T, Gerdts V, Perez-Casal J. 2011. Protein chimeras containing the Mycoplasma bovis GAPDH protein and bovine host-defence peptides retain the properties of the individual components. Microbial Pathogenesis, 50, 269–277.
Minst K, Martlbauer E, Miller T, Meyer C. 2012. Short communication: Streptococcus species isolated from mastitis milk samples in Germany and their resistance to antimicrobial agents. Journal of Dairy Science, 95, 6957–6962.
Neiwert O, Holst O, Duda K A. 2014. Structural investigation of rhamnose-rich polysaccharides from Streptococcus dysgalactiae bovine mastitis isolate. Carbohydrate Research, 389, 192–195.
Oliver S P, Pighetti G M. 2002. Mastitis Pathogens: Environmental pathogens. Encyclopedia of Dairy Sciences, 3, 1728–1734.
Park Y K, Nho S W, Shin G W, Park S B, Jang H B, Cha I S, Ha M A, Kim Y R, Dalvi R S, Kang B J, Jung T S. 2009. Antibiotic susceptibility and resistance of Streptococcus iniae and Streptococcus parauberis isolated from olive flounder (Paralichthys olivaceus). Veterinary Microbiology, 136, 76–81.
Perrig M S, Ambroggio M B, Buzzola F R, Marcipar I S, Calvinho L F, Veaute C M, Barbagelata M S. 2015. Genotyping and study of the pauA and sua genes of Streptococcus uberis isolates from bovine mastitis. Revista Argentina de Microbiología, 47, 282–294.
Pitkala A, Koort J, Bjorkroth J. 2008. Identification and antimicrobial resistance of Streptococcus uberis and Streptococcus parauberis isolated from bovine milk samples. Journal of Dairy Science, 91, 4075–4081.
Poutrel B, Bareille S, Lequeux G, Leboeuf F. 2018. Prevalence of mastitis pathogens in France: Antimicrobial susceptibility of Staphylococcus aureus, Streptococcus uberis and Escherichia coli. Journal of Veterinary Science & Technology, 9, 522.
Rato M G, Bexiga R, Florindo C, Cavaco L M, Vilela C L, Santos-Sanches I. 2013. Antimicrobial resistance and molecular epidemiology of streptococci from bovine mastitis. Veterinary Microbiology, 161, 286–294.
Reinoso E B, Lasagno M C, Dieser S A, Odierno L M. 2011. Distribution of virulence-associated genes in Streptococcus uberis isolated from bovine mastitis. FEMS Microbiology Letters, 318, 183–188.
Roy S, Gao G, Lu Y, Zhou X, Lock M, Calcedo R, Wilson J M. 2004. Characterization of a family of Chimpanzee adenoviruses and development of molecular clones for gene transfer vectors. Human Gene Therapy, 15, 519–530.
Ruegg P L. 2012. Managing Cows, milking and the environment to minimize mastitis. Advances in Dairy Technology, 24, 351–359.
Savini V, Marrollo R, Coclite E, Fusilli P, D’incecco C, Fazii P, Gherardi G. 2014. Liofilchem® Chromatic VRE and vancomycin MIC Test Strip detected glycopeptide resistance in a vanB neonatal Enterococcus faecium isolate showing alternate vancomycin susceptibility and resistance with bioMérieux Vitek2. International Journal of Clinical and Experimental Pathology, 7, 6274–6277.
Schwarz D, Diesterbeck U S, Konig S, Brugemann K, Schlez K, Zschock M, Wolter W, Czerny C P. 2011. Microscopic differential cell counts in milk for the evaluation of inflammatory reactions in clinically healthy and subclinically infected bovine mammary glands. Journal of Dairy Research, 78, 448–455.
Song B, Yang X, Sun H, Yu L, Ma J, Wu Z, Cui Y. 2017. Immunogenicity of amino acids 1–150 of Streptococcus GapC displayed on the surface of Escherichia coli. Microbial Pathogenesis, 105, 288–297.
Stevens M, Piepers S, De Vliegher S. 2016. Mastitis prevention and control practices and mastitis treatment strategies associated with the consumption of (critically important) antimicrobials on dairy herds in Flanders, Belgium. Journal of Dairy Science, 99, 2896–2903.
Takaya A, Kitagawa N, Kuroe Y, Endo K, Okazaki M, Yokoyama E, Wada A, Yamamoto T. 2010. Mutational analysis of reduced telithromycin susceptibility of Streptococcus pneumoniae isolated clinically in Japan. FEMS Microbiology Letters, 307, 87–93.
Thomas V, De Jong A, Moyaert H, Simjee S, El Garch F, Morrissey I, Marion H, Valle M. 2015. Antimicrobial susceptibility monitoring of mastitis pathogens isolated from acute cases of clinical mastitis in dairy cows across Europe: VetPath results. International Journal of Antimicrobial Agents, 46, 13–20.
Ward P N, Leigh J A. 2004. Genetic analysis of Streptococcus uberis plasminogen activators. The Indian Journal of Medical Research, 119, 136–140.
Williams A M, Collins M D. 1990. Molecular taxonomic studies on Streptococcus uberis types I and II. Description of Streptococcus parauberis sp. nov. The Journal of applied bacteriology, 68, 485–490.
Yan Y, Hu H, Lu T, Fan H, Hu Y, Li G, Zhang X, Shi Y, Xia R. 2016. Investigation of serotype distribution and resistance genes profile in group B Streptococcus isolated from pregnant women: A Chinese multicenter cohort study. APMIS, 124, 794–799.
Yang F, Wang Q, Wang X R, Wang L, Li X P, Luo J Y, Zhang S D, Li H S. 2016. Genetic characterization of antimicrobial resistance in Staphylococcus aureus isolated from bovine mastitis cases in Northwest China. Journal of Integrative Agriculture, 15, 2842–2847.
Yang F, Zhang S, Shang X, Wang X, Yan Z, Li H, Li J. 2019. Short communication: Antimicrobial resistance and virulence genes of Enterococcus faecalis isolated from subclinical bovine mastitis cases in China. Journal of Dairy Science, 102, 140–144.
Zhong C Y, Cheng A C, Wang M S, Zhu D K, Luo Q H, Chen S, Zhang S H, Chen X Y. 2013. Quantitative real-time PCR study of the expression and regulation of the tetracycline resistance gene in Riemerella anatipestifer. Poultry Science, 92, 1552–1559.
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