Methylene blue-mediated antimicrobial photodynamic therapy on chicken semen

Background: Artificial insemination is widely employed in poultry, but high degrees of bacterial contamination are often observed in semen because of its passage through the cloaca. Consequently, most semen extenders for birds have antibiotics that could aggravate bacterial resistance.Methods: We evaluated the potential of antimicrobial photodynamic therapy (PDT) as an alternative to the use of antibiotics, and assessed whether changes in concentration and incubation time with methylene blue (MB), radiant exposure, and irradiance of light affect spermatozoa activity and bacteria in chicken semen.Results: Incubation with MB (< 25 mu M) did not alter sperm motility, regardless of the pre-irradiation time (PIT, 1 or 5 min). Following 1 min of PIT with MB at 10 mu M, samples were irradiated for 30, 60, 120, and 180 s at irradiances of 44, 29, and 17 mW/ cm2 (660 nm LedBox). MB and light alone did not interfere with the analyzed parameters. However, when both factors were associated, increases in light dose led to greater reductions in sperm parameters, regardless of the irradiance used. Besides, PDT conditions that were less harmful to sper-matozoa were not able to significantly reduce bacterial colonies in chicken semen.Conclusions: A failure in MB selectivity could explain unsuccessful bacterial reduction following PDT. Further research involving other photosensitizers or conjugating molecules to MB to target microbial cells is needed for PDT application in poultry breeders.

Automated summary

Artificial insemination in poultry often leads to high levels of bacterial contamination in the semen. This study explored the potential use of antimicrobial photodynamic therapy (PDT) as an alternative to antibiotics in preserving semen quality. The results showed that incubation with methylene blue (MB) did not affect sperm motility, but increasing light dose during the PDT process resulted in greater reductions in sperm parameters. However, the conditions that were less harmful to spermatozoa were not effective in reducing bacterial colonies in chicken semen.