Short communication: Barrier characteristics of 3 external teat sealants to prevent bacterial penetration under in vitro conditions using rubber calf -feeding nipples

Abstract
The main objective of this study was to evaluate the barrier characteristics of 3 external teat sealants for dry cows in preventing bacterial penetration by 3 common major mastitis pathogens (Escherichia coli, Staphylococcus aureus, and Streptococcus uberis) via a novel in vitro simulation model using rubber calf-feeding nipples. All feeding nipples were filled with a sterile cotton plug soaked in sterile broth heart infusion medium and were treated as follows: rubber teats 1 and 5 were sealed with Ubera Dry (Inovet, Arendonk, Belgium); rubber teats 2 and 6 were sealed with T-Hexx Dry (Huvepharma Livestock, St. Louis, MO); rubber teats 3 and 7 were sealed with Uddergold Dry (Ecolab Food and Beverage Division, St. Paul, MN); and rubber teats 4 and 8 remained unsealed and served as positive and negative controls, respectively. After the dips had dried, rubber teats 1 to 4 were immersed in a suspension of E. coli (experiment 1), Staph. aureus (experiment 2), or Strep. uberis (experiment 3; ≥1.5 × 108 cfu/mL) for 24 h, whereas rubber teats 5 to 8 were not exposed to the bacterial suspensions. All external teat sealants adhered well to the rubber teats. All cotton plugs collected from the teats that were not exposed to E. coli, Staph. aureus, or Strep. uberis (rubber teats 5 to 8) remained culture-negative, except for 1 (due to contamination). Of the rubber teats that were exposed to the major mastitis pathogens, all cotton plugs collected from the teats dipped with Ubera Dry and T-Hexx Dry remained culture-negative for the mastitis pathogen they were exposed to. The cotton plugs of the rubber teats that were sealed with Uddergold Dry and exposed to E. coli and Strep. uberis showed positive cultures for the respective bacteria, as demonstrated using strain-typing. The cotton plugs collected from the rubber teats that were not sealed with an external teat sealant and that served as positive controls became culture-positive for the mastitis pathogens they were exposed to. We conclude that Ubera Dry showed comparable and superior barrier performance against penetration of E. coli, Staph. aureus, and Strep. uberis compared with T-Hexx Dry and Uddergold Dry, respectively, under in vitro conditions using a novel in vitro simulation model. Although one should be aware that the method has not yet been validated to predict risk of intramammary infections, the proposed technique can be a meaningful starting point to evaluate and compare the barrier characteristics of external teat sealants in preventing bacterial penetration. A large-scale clinical trial is needed before any definite conclusions can be drawn as to the adherence, duration of adherence, barrier performance, and efficacy in protection against intramammary infections of the 3 external teat sealants under field conditions.

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