In vitro impact of macrolide antibiotics on the viability of selected mammalian cell lines

Eva Tvrdá


The aim of this study was to evaluate the in vitro cytotoxicity of different concentrations of macrolide  antibiotics (tilmicosin-TILM, tylosin-TYL and spiramycin-SPI) on selected animal cell cultures. VERO cells (kidney cells from Macacus rhesus), FE cells (feline embryonal cells) and BHK 21 cells (cell line from young hamster kidneys) were used in the study and subjected to concentrations of macrolides ranging 50-1000 µg/mL, depending on the specific antibiotic and cell line. The cell viability expressed as the mitochondrial activity of living cells was assessed using the metabolic mitochondrial MTT test. The effect of tilmicosin: FE cells were the most sensitive with a significant decrease of mitochondrial activity at 100-150 µg/mL (P<0.001) TILM. VERO cells were the most resistant, as no significant decrease of viability was observed at any TILM dose. The effect of tylosin: FE cells showed the highest sensitivity to TYL, as 1000 µg/mL reduced the cell viability to a half (P<0.001) when compared to the untreated control. Similarly, a decreased viability of BHK 21 cells was observed following the supplementation of 1500 (P<0.001) and 900 (P<0.05) µg/mL TYL. VERO cells exhibited the highest resilience to the TYL treatment, with no significant differences of viability in comparison to the control. The effect of spiramycin: BHK 21 cells exhibited the highest sensitivity to the antibiotic, as all concentrations (150, 200, 300 µg/mL SPI) led to a significant decrease (P<0.001) of the mitochondrial activity.  Similarly, the viability of FE cells significantly (P<0.05) decreased after the administration of 350 and 540 µg/mL SPI. On the other hand, VERO cells revealed the highest resistance to the antibiotic, with no significant effects in comparison to the control. Our data reveal that macrolides have a significant adverse negative effect on the cell viability, and may provide more information to our knowledge on the specific effects medication has on the organism.


cell cultures; macrolides; mitochondrial activity; spiramycin; tilmicosin; tylosin; viability

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Nikolaou, A., Meric, S., Fatta, D., Occurrence patterns of pharmaceuticals in water and wastewater environments. Analytical and Bioanalytical Chemistry, 2007, 387, 1225-1234.

Lai, H. T., Hou, J. H., Su, C. I., Chen, C. L., Effects of chloramphenicol, florfenicol, and thiamphenicol on growth of algae Chlorella pyrenoidosa, Isochrysis galbana, and Tetraselmis chui. Ecotoxicology and Environmental Safety, 2009, 72, 329-734.

Kümmerer, K., Antibiotics in the aquatic environment-a review-part II. Chemosphere, 2009, 75, 435-441.

Baran, W., Adamek, E., Ziemiańska, J., Sobczak, A., Effects of the presence of sulfonamides in the environment and their influence on human health. Journal of Hazardous Materials, 2011, 196, 1-15.

Giguere, S., Prescott, J.F., Baggot, J.D., Walker, R.D., Dowling, P.M., Antimicrobial Therapy in Veterinary Medicine, fourth ed., Wiley-Blackwell, New York, 2007, 626 p.

Hamilton-Miller, J. M., Chemistry and biology of the polyene macrolide antibiotics. Bacteriological Reviews, 1973, 37, 166-196.

Kanoh, S., Rubin, B. K., Mechanisms of action and clinical application of macrolides as immunomodulatory medications. Clinical Microbiology Reviews, 2010, 23, 590-615.

De Liguoro, M., Fioretto, B., Poltronieri, C., Gallina, G., The toxicity of sulfamethazine to Daphnia magna and its additivity to other veterinary sulfonamides and trimethoprim. Chemosphere, 2009, 75, 1519-1524.

Eguchi, K., Nagase, H., Ozawa, M., Endoh, Y. S., Goto, K., Hirata, K., Miyamoto, K., Yoshimura, H., Evaluation of antimicrobial agents for veterinary use in the ecotoxicity test using microalgae. Chemosphere, 2004, 57, 1733-1738.

Isidori, M., Lavorgna, M., Nardelli, A., Pascarella, L., Parrella, A., Toxic and genotoxic evaluation of six antibiotics on non-target organisms. Science of The Total Environment, 2005, 346, 87-98.

Lin, A.Y.C., Yu, T.H., Lin, C.F., Pharmaceutical contamination in residential, industrial, and agricultural waste streams: risk to aqueous environments in Taiwan, Chemosphere, 2008, 74, 131-141.

Tvrdá, E., Lukáč, N., Lukáčová, J., Jambor, T., Massányi, P., Dose- and time-dependent in vitro effects of divalent and trivalent iron on the activity of bovine spermatozoa. Biological Trace Element Research, 2015, 167, 36-47.

Shryock, T.R., Staples, J.M., DeRosa, D.C., Minimum inhibitory concentration breakpoints and disk diffusion inhibitory zone interpretive criteria for tilmicosin susceptibility testing against Pasteurella multocida and Actinobacillus pleuropneumoniae associated with porcine respiratory disease. Journal of Veterinary Diagnostic Investigation, 2000, 14, 389-395.

Ishida, Y., Abe, Y., Harabuchi, Y., Effects of macrolides on antigen presentation and cytokine production by dendritic cells and T lymphocytes. International Journal of Pediatric Otorhinolaryngology, 2007, 71, 397-405.

Viluksela, M., Vainio, P. J., Tuominen, R. K., Cytotoxicity of macrolide antibiotics in a cultured human liver cell line. Journal of Antimicrobial Chemothrapy, 1996, 38, 465-473.

Otoguro, K., Komiyama, K., Omura, S., Tyson, C.A., An in vitro cytotoxicity assay using rat hepatocytes and MTT and Coomassie blue dye as indicators. Alternatives to Laboratory Animals, 1991, 19, 352-360.

Jordan, W. H., The toxicity of tilmicosin given orally to rats for three months, R-09886. Lilly research Laboratories, Toxicological Division, 1989. Web page:

Anderson, R. C., Harris, P. N., Lee, C. C., Maze, N., Small, R. M., Worth, H. M., The toxicology and pharmacology of tylosin, an antibiotic, and some salts of tylosin, X/E/4. Lilly research Laboratories, Toxicological Division, 1966. Web page:

Baguer, A. J., Jensen, J., Krogh, P. H., Effects of the antibiotics oxytetracycline and tylosin on soil fauna. Chemosphere, 2000, 40, 751-757.

The European Agency for the Evaluation of Medicinal Products, Veterinary Medicines Evaluation Unit, 1994. Web page:

Havlík, J., Význam makrolidových antibiotik. Lékařské listy, 2008, 5, 7-9.


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