FREE SHIPPING over $40
OVER 9000 FIVE-STAR REVIEWS
5% OFF ALL SUBSCRIPTIONS
100% HAPPINESS GUARANTEE
Analyze Diet
0
Home / Equine Research Bank / Reprod Domest Anim / Penicillin and amikacin mixture has bactericidal activity eq...
Reproduction in domestic animals = Zuchthygiene2022; 58(2); 349-357; doi: 10.1111/rda.14294

Penicillin and amikacin mixture has bactericidal activity equivalent to gentamicin, tylosin, lincomycin and spectinomycin mixture in equine frozen semen.

Abstract: Neat stallion semen can contain a variety of microorganisms, some of which may impair sperm quality and/or cause infection of the mares' reproductive tract. For this reason, antibiotics are commonly added to semen extenders. A combination of gentamicin, tylosin, lincomycin and spectinomycin (GTLS) has been recommended for use, but there are no reports on the use of this mixture in equine semen extender. Penicillin and amikacin (PA) are safe for preserving sperm quality while effectively controlling bacterial growth in equine cooled stored semen, but data on frozen semen are scarce. Therefore, a bioequivalence study was performed to assess the bactericidal activity of GTLS and PA in equine frozen semen. Nine mature, healthy stallions were used in the study. Split ejaculates were processed using media without antibiotics (Control) or with different antibiotics. For the GTLS group, centrifugation medium and freezing extender were prepared with gentamicin 250 μg/ml, tylosin 50 μg/ml, lincomycin 150 μg/ml and spectinomycin 300 μg/ml. For the PA group, the centrifugation medium was prepared with potassium penicillin G (PPG) 1200 units/ml and the freezing extender was prepared with PPG 1200 units/ml and amikacin 500 μg/ml. Semen processed in extenders without antibiotics had higher (p < .005) bacterial loads throughout all cryopreservation processing steps than semen samples processed using antibiotics. There were no differences in semen bacterial load after centrifugation, 15 and 30 min after final extension, and after thawing between GTLS and PA groups, but PA had faster (p < .05) kill-time kinetics than GTLS. Only minor differences in sperm kinetic parameters were observed among groups. In conclusion, this study demonstrated bioequivalence between GTLS and PA in mitigating end-point bacterial loads. Prudent concentrations of the antibiotic mixtures evaluated in this study can be considered both effective and sperm-safe for equine frozen semen.
© 2022 Wiley-VCH GmbH.
Get Full Text
Publication Date: 2022-11-25 PubMed ID: 36369673DOI: 10.1111/rda.14294Google Scholar: Lookup
The Equine Research Bank provides access to a large database of publicly available scientific literature. Inclusion in the Research Bank does not imply endorsement of study methods or findings by Mad Barn.
LinkedInCopy
  • Journal Article

Summary

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

The research paper has studied the effectiveness of the Penicillin and Amikacin (PA) mixture and Gentamicin, Tylosin, Lincomycin and Spectinomycin (GTLS) mixture in protecting equine frozen semen from bacterial growth. The researchers have found that both mixtures are equally effective in controlling sperm-safe bacteria in horse’s frozen semen.

Introduction

The research was initiated to assess the bactericidal activity of two different antibiotic mixtures – Penicillin and Amikacin (PA) and Gentamicin, Tylosin, Lincomycin and Spectinomycin (GTLS) in equine frozen semen. It was based on the background that stallion semen can contain numerous microorganisms which might impair sperm quality or cause infection in the mares’ reproductive tract. To curb this, antibiotics are often added to semen extenders.

Methodology

  • Nine healthy, mature stallions were used for the study and split ejaculates were processed using media with varied antibiotics.
  • The GTLS mixture consisted of gentamicin 250 μg/ml, tylosin 50 μg/ml, lincomycin 150 μg/ml, and spectinomycin 300 μg/ml while the PA mixture was prepared with potassium penicillin G (PPG) 1200 units/ml and amikacin 500 μg/ml added in the freezing extender along with PPG 1200 units/ml in the centrifugation medium.

Results

  • The researchers found that semen treated with extenders without the antibiotics showed greater bacterial loads during all cryopreservation procedures than those treated with antibiotics.
  • Bacterial load in the semen after centrifugation, and after 15 and 30 minutes of final extension and thawing, showed no significant difference between GTLS and PA groups.
  • Interestingly, PA had faster kill-time kinetics than GTLS. However, only slight variations were seen in the sperm kinetic parameters across groups.

Conclusion

In conclusion, it was observed that both GTLS and PA are bioequivelant in mitigating end-point bacterial loads. Therefore, it suggests that these antibiotic mixtures can be considered effective and safe for preserving equine frozen semen.

Cite This Article

APA
Brito LFC, Loomis PR, Klohonatz KM, Althouse GC. (2022). Penicillin and amikacin mixture has bactericidal activity equivalent to gentamicin, tylosin, lincomycin and spectinomycin mixture in equine frozen semen. Reprod Domest Anim, 58(2), 349-357. https://doi.org/10.1111/rda.14294

Publication

Reproduction in domestic animals = Zuchthygiene
ISSN: 1439-0531
NlmUniqueID: 9015668
Country: Germany
Language: English
Volume: 58
Issue: 2
Pages: 349-357

Researcher Affiliations

Brito, Leonardo F C
  • Department of Clinical Studies - New Bolton Center, School of Veterinary Medicine, University of Pennsylvania, Pennsylvania, Kennett, USA.
Loomis, Paul R
  • Select Breeders Service, Chesapeake, Maryland, USA.
Klohonatz, Kristin M
  • Select Breeders Service, Chesapeake, Maryland, USA.
Althouse, Gary C
  • Department of Clinical Studies - New Bolton Center, School of Veterinary Medicine, University of Pennsylvania, Pennsylvania, Kennett, USA.

MeSH Terms

  • Animals
  • Horses
  • Male
  • Female
  • Spectinomycin / pharmacology
  • Lincomycin / pharmacology
  • Tylosin
  • Amikacin / pharmacology
  • Gentamicins / pharmacology
  • Penicillins
  • Semen Preservation / veterinary
  • Semen / microbiology
  • Anti-Bacterial Agents / pharmacology
  • Spermatozoa / microbiology
  • Cryopreservation / veterinary
  • Sperm Motility

Grant Funding

  • Minitube
  • Select Breeders Service

References

This article includes 34 references
  1. Al-Kass Z, Eriksson E, Bagge E, Wallgren M, Morrell JM. Microbiota of semen from stallions in Sweden identified by MALDI-TOF. Veterinary and Animal Science 2020;10:100143.
    doi: 10.1016/j.vas.2020.100143google scholar: lookup
  2. Al-Kass Z, Spergser J, Aurich C, Kuhl J, Schmidt K, Johannisson A, Morrell JM. Sperm quality during storage is not affected by the presence of antibiotics in EquiPlus semen extender but is improved by single layer centrifugation. Antibiotics 2017;7(1):1-13.
    doi: 10.3390/antibiotics7010001google scholar: lookup
  3. Al-Kass Z, Spergser J, Aurich C, Kuhl J, Schmidt K, Morrell JM. Effect of presence or absence of antibiotics and use of modified single layer centrifugation on bacteria in pony stallion semen. Reproduction in Domestic Animals 2019;54(2):342-349.
    doi: 10.1111/rda.13366google scholar: lookup
  4. Althouse G, Skaife J, Loomis P. Prevalence and types of contaminant bacteria in extended, chilled equine semen. Animal Reproduction Science 2010;121S:S224-S225.
    doi: 10.13140/rg.2.1.4670.4808google scholar: lookup
  5. Alvarenga MA, Papa FO, Ramires Neto C. Advances in stallion semen cryopreservation. The Veterinary Clinics of North America. Equine Practice 2016;32(3):521-530.
    doi: 10.1016/j.cveq.2016.08.003google scholar: lookup
  6. Arriola J, Foote RH. Effects of amikacin sulfate on the motility of stallion and bull spermatozoa at different temperatures and intervals of storage. Journal of Animal Science 1982;54(6):1105-1110.
    doi: 10.2527/jas1982.5461105xgoogle scholar: lookup
  7. Aurich C, Spergser J. Influence of bacteria and gentamicin on cooled-stored stallion spermatozoa. Theriogenology 2007;67(5):912-918.
    doi: 10.1016/j.theriogenology.2006.11.004google scholar: lookup
  8. Breuil MF, Duquesne F, Laugier C, Petry S. Phenotypic and 16 S ribosomal RNA gene diversity of Taylorella asinigenitalis strains isolated between 1995 and 2008. Veterinary Microbiology 2011;148(2-4):260-266.
    doi: 10.1016/j.vetmic.2010.08.028google scholar: lookup
  9. Corona A, Cherchi R. Microbial quality of equine frozen semen. Animal Reproduction Science 2009;115(1-4):103-109.
    doi: 10.1016/j.anireprosci.2008.11.016google scholar: lookup
  10. CSS minimum requirements for disease control of semen produced for artificial insemination, (2021). https://www.naab-css.org/uploads/userfiles/files/20220720%20CSSMinReq%20Jan2021-ENG_Update_Final_v_2.pdf
  11. Delerue M, Breuil MF, Duquesne F, Bayon-Auboyer MH, Amenna-Bernard N, Petry S. Acute Endometritis due to Taylorella equigenitalis transmission by insemination of cryopreserved stallion semen. Journal of Equine Veterinary Science 2019;78:10-13.
    doi: 10.1016/j.jevs.2019.03.217google scholar: lookup
  12. EU Commision Delegated Regulation (EU) 2020/686 supplementing Regulation (EU) 2016/429 of the European Parliament and of the Council as regards the approval of germinal product establishments and the traceability and animal health requirements for movements within the Union of germinal products of certain kept terrestrial animals. (2019). https://eur-lex.europa.eu/legal-content/EN/TXT/PDF/?uri=CELEX:32020R0686&from=EN
  13. Guimarães T, Lopes G, Pinto M, Silva E, Miranda C, Correia MJ, Damásio L, Thompson G, Rocha A. Colloid centrifugation of fresh stallion semen before cryopreservation decreased microorganism load of frozen-thawed semen without affecting seminal kinetics. Theriogenology 2015;83(2):186-191.
    doi: 10.1016/j.theriogenology.2014.09.003google scholar: lookup
  14. Guimarães T, Miranda C, Pinto M, Silva E, Damásio L, Costa AL, Correia MJ, Duarte JC, Cosinha C, Lopes G, Thompson G, Rocha A. Effect of breeding activity on the microflora of the external genitalia and in the semen of stallions, and the relationship between micro-organisms on the skin and on the external genitalia. Reproduction in Domestic Animals 2014;49(6):926-933.
    doi: 10.1111/rda.12403google scholar: lookup
  15. Hernandez-Aviles C, Love CC, Serafini R, Ramirez-Agamez L, Kelley DE, de Andino EM, Teague SR, LaCaze KA, Brinsko SP, Varner DD. Inclusion of supplemental antibiotics (amikacin - penicillin) in a commercial extender for stallion semen: Effects on sperm quality, bacterial growth, and fertility following cooled storage. Theriogenology 2020;158:209-217.
    doi: 10.1016/j.theriogenology.2020.09.018google scholar: lookup
  16. Hernandez-Aviles C, Serafini R, Love CC, Teague SR, LaCaze KA, Lawhon SD, Wu J, Blanchard TL, Varner DD. The effects of antibiotic type and extender storage method on sperm quality and antibacterial effectiveness in fresh and cooled-stored stallion semen. Theriogenology 2018;122:23-29.
    doi: 10.1016/j.theriogenology.2018.08.022google scholar: lookup
  17. Jasko DJ, Bedford SJ, Cook NL, Mumford EL, Squires EL, Pickett BW. Effect of antibiotics on motion characteristics of cooled stallion spermatozoa. Theriogenology 1993;40(5):885-893.
    doi: 10.1016/0093-691x(93)90356-agoogle scholar: lookup
  18. Klein C, Donahue JM, Sells SF, Squires EL, Timoney PJ, Troedsson MH. Effect of antimicrobial-containing semen extender on risk of dissemination of contagious equine metritis. Journal of the American Veterinary Medical Association 2012;241(7):916-921.
    doi: 10.2460/javma.241.7.916google scholar: lookup
  19. Lorton SP, Sullivan JJ, Bean B, Kaproth M, Kellgren H, Marshall C. A new antibiotic combination for frozen bovine semen, 3. Evaluation of fertility. Theriogenology 1988;29(3):609-614.
    doi: 10.1016/s0093-691x(88)80008-6google scholar: lookup
  20. Lorton SP, Sullivan JJ, Bean B, Kaproth M, Kellgren H, Marshall C. A new antibiotic combination for frozen bovine semen. 2. Evaluation of seminal quality. Theriogenology 1988;29(3):593-607.
    doi: 10.1016/s0093-691x(88)80007-4google scholar: lookup
  21. Malmgren L, Olsson Engvall E, Engvall A, Albihn A. Aerobic bacterial flora of semen and stallion reproductive tract and its relation to fertility under field conditions. Acta Veterinaria Scandinavica 1998;39(2):173-182.
    doi: 10.1186/bf03547790google scholar: lookup
  22. Olivieri B, Love B, Rezabek G, Lamm C, Varner D, Payton M, Holyoak G. Effect of antibiotic-containing extenders on Taylorella equigenitalis contaminated semen. Journal of Equine Veterinary Science 2011;31:655-660.
    doi: 10.1016/j.jevs.2011.04.002google scholar: lookup
  23. Ortega-Ferrusola C, González-Fernández L, Muriel A, Macías-García B, Rodríguez-Martínez H, Tapia JA, Alonso JM, Peña FJ. Does the microbial flora in the ejaculate affect the freezeability of stallion sperm?. Reproduction in Domestic Animals 2009;44(3):518-522.
    doi: 10.1111/j.1439-0531.2008.01267.xgoogle scholar: lookup
  24. Papich MG. Beta-lactam antibiotics: Penicillins, cephalosporins, and related drugs. .
  25. Papich MG, Riviere JE. Aminoglycoside antibiotics. .
  26. Pasing SS, Aurich C, von Lewinski M, Wulf M, Krüger M, Aurich JE. Development of the genital microflora in stallions used for artificial insemination throughout the breeding season. Animal Reproduction Science 2013;139(1-4):53-61.
    doi: 10.1016/j.anireprosci.2013.03.009google scholar: lookup
  27. Price S, Aurich J, Davies-Morel M, Aurich C. Effects of oxygen exposure and gentamicin on stallion semen stored at 5 and 15 degrees C. Reproduction in Domestic Animals 2008;43(3):261-266.
    doi: 10.1111/j.1439-0531.2007.00888.xgoogle scholar: lookup
  28. Rota A, Calicchio E, Nardoni S, Fratini F, Ebani VV, Sgorbini M, Panzani D, Camillo F, Mancianti F. Presence and distribution of fungi and bacteria in the reproductive tract of healthy stallions. Theriogenology 2011;76(3):464-470.
    doi: 10.1016/j.theriogenology.2011.02.023google scholar: lookup
  29. Samper JC, Tibary A. Disease transmission in horses. Theriogenology 2006;66(3):551-559.
    doi: 10.1016/j.theriogenology.2006.04.019google scholar: lookup
  30. Shin SJ, Lein DH, Patten VH, Ruhnke HL. A new antibiotic combination for frozen bovine semen 1. Control of mycoplasmas, ureaplasmas, campylobacter fetus subsp. venerealis and Haemophilus somnus. Theriogenology 1988;29(3):577-591.
    doi: 10.1016/s0093-691x(88)80006-2google scholar: lookup
  31. USDA-APHIS Model Animal Health Certificate for entry into the union of consignments of semen of equine animals collected, processed and stored in accordance with Regulation (EU) 2016/429 and delegated regulation (EU) 2020/692 after April 20, 2021., (2021). https://www.aphis.usda.gov/regulations/vs/iregs/animals/downloads/eu-equi-sem-a-entry-hc.pdf
  32. Varela E, Rey J, Plaza E, Muñoz de Propios P, Ortiz-Rodríguez JM, Álvarez M, Anel-López L, Anel L, De Paz P, Gil MC, Morrell JM, Ortega-Ferrusola C. How does the microbial load affect the quality of equine cool-stored semen?. Theriogenology 2018;114:212-220.
    doi: 10.1016/j.theriogenology.2018.03.028google scholar: lookup
  33. Varner DD, Scanlan CM, Thompson JA, Brumbaugh GW, Blanchard TL, Carlton CM, Johnson L. Bacteriology of preserved stallion semen and antibiotics in semen extenders. Theriogenology 1998;50(4):559-573.
    doi: 10.1016/s0093-691x(98)00161-7google scholar: lookup
  34. Wilsher S, Omar H, Ismer A, Allen T, Wernery U, Joseph M, Mawhinney I, Florea L, Thurston L, Duquesne F, Petry S. A new strain of Taylorella asinigenitalis shows differing pathogenicity in mares and Jenny donkeys. Equine Veterinary Journal 2021;53(5):990-995.
    doi: 10.1111/evj.13382google scholar: lookup
Subscribe to our newsletter
Join 99,344 horse owners like you who receive our email updates on horse health and nutrition.