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Synergistic streptococcal phage λSA2 and B30 endolysins kill streptococci in cow milk and in a mouse model of mastitis

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Abstract

Bovine mastitis results in billion dollar losses annually in the USA alone. Streptococci are among the most relevant causative agents of this disease. Conventional antibiotic therapy is often unsuccessful and contributes to development of antibiotic resistance. Bacteriophage endolysins represent a new class of antimicrobials against these bacteria. In this work, we characterized the endolysins (lysins) of the streptococcal phages λSA2 and B30 and evaluated their potential as anti-mastitis agents. When tested in vitro against live streptococci, both enzymes exhibited near-optimum lytic activities at ionic strengths, pH, and Ca2+ concentrations consistent with cow milk. When tested in combination in a checkerboard assay, the lysins were found to exhibit strong synergy. The λSA2 lysin displayed high activity in milk against Streptococcus dysgalactiae (reduction of CFU/ml by 3.5 log units at 100 μg/ml), Streptococcus agalactiae (2 log), and Streptococcus uberis (4 log), whereas the B30 lysin was less effective. In a mouse model of bovine mastitis, both enzymes significantly reduced intramammary concentrations of all three streptococcal species (except for B30 vs. S. dysgalactiae), and the effects on mammary gland wet weights and TNFα concentrations were consistent with these findings. Unexpectedly, the synergistic effect determined for the two enzymes in vitro was not observed in the mouse model. Overall, our results illustrate the potential of endolysins for treatment of Streptococcus-induced bovine mastitis.

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Acknowledgments

Mentioning of trade names or commercial products in this article is solely for the purpose of providing specific information and does not imply recommendation or endorsement by the US Department of Agriculture (USDA). The USDA prohibits discrimination in all its programs and activities on the basis of race, color, national origin, age, disability, and where applicable, sex, marital status, familial status, parental status, religion, sexual orientation, genetic information, political beliefs, reprisal, or because all or part of an individual’s income is derived from any public assistance program. USDA is an equal opportunity provider and employer.

Funding

This work was supported in part by National Institutes of Health grant 1 R01 AI075077-01A1; National Research Initiative grant 2007-35204-18395 and US State Department funds supporting US-Pakistani and US-Russian collaborations.

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ᅟ

Conflict of interest

The authors declare that they have no conflict of interest.

Statement regarding research on animals

All animal experiments were conducted in accordance with ARS Institutional Animal Care and Use Committee regulations and national animal care guidelines.

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  1. Calvin S. Pohl

    Present address: North Carolina State College of Veterinary Medicine, 1060 William Moore Drive, Raleigh, NC, 27607, USA

Authors and Affiliations

  1. Animal Biosciences and Biotechnology Laboratory, North East Area, Agricultural Research Service, U.S. Department of Agriculture, 10300 Baltimore Ave, Beltsville, MD, 20705, USA

    Mathias Schmelcher, Anne M. Powell, Calvin S. Pohl & David M. Donovan

  2. Institute of Food, Nutrition and Health, ETH Zurich, Schmelzbergstrasse 7, LFV B18, 8092, Zurich, Switzerland

    Mathias Schmelcher

  3. Statistics Group, Office of the North East Area Director, Agricultural Research Service, U.S. Department of Agriculture, 10300 Baltimore Ave, Beltsville, MD, 20705, USA

    Mary J. Camp

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Correspondence to Mathias Schmelcher.

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Schmelcher, M., Powell, A.M., Camp, M.J. et al. Synergistic streptococcal phage λSA2 and B30 endolysins kill streptococci in cow milk and in a mouse model of mastitis. Appl Microbiol Biotechnol 99, 8475–8486 (2015). https://doi.org/10.1007/s00253-015-6579-0

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