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The bovine oviduct provides the site for fertilization and early embryonic development. Modifications to
this physiological environment, for instance the presence of pathogenic bacterial species, could diminish
reproductive success at early stages of pregnancy. The aim of this study was to elucidate the inflammatory
responses of bovine oviductal epithelial cells (BOEC) to a pathogenic bacterial species (Trueperella
pyogenes) and a potentially pathogenic bacterium (Bacillus pumilus). BOEC from four healthy animals
were isolated, cultured in passage 0 (P0) and passaged until P3. Trypan blue staining determined BOEC
viability during 24 h co-culture with different multiplicities of infection (MOI) of T. pyogenes (MOI 0.01,
0.05, 0.1 and 1) or B. pumilus (MOI 1 and 10). BOEC remained viable when co-cultured with T. pyogenes at
MOI 0.01 and with B. pumilus at MOI 1 and 10. Extracted total RNA from control and bacteria co-cultured
samples was subjected to reverse transcription-quantitative polymerase chain reaction (RTq-PCR) to
determine mRNA expression of various studied genes. The rate of release of interleukin 8 (IL8) and
prostaglandin E2 (PGE2) from BOEC was measured by ELISA after 24 h co-culture with bacteria. RT-qPCR
of various selected pro-inflammatory factors revealed similar mRNA expression of pro-inflammatory
factors in BOEC co-cultured with T. pyogenes and in the controls. Higher mRNA expression of IL 1A,
-1B, tumor necrosis factor alpha and CXC ligand (CXCL) 1/2, 3, 5 and IL8 and PG synthesis enzymes in
BOEC co-cultured with B. pumilus was observed. In the presence of B. pumilus a higher amount of IL8 and
PGE2 was released from BOEC than from controls. The viability and pro-inflammatory response of P3
BOEC incubated with bacteria was lower than in P0 BOEC. These findings illustrate the pathogenicity of
T. pyogenes towards BOEC in detail and the potential role of B. pumilus in generating inflammation in
oviductal cells. Culturing conditions influenced the pro-inflammatory responses of BOEC towards bacteria.
Therefore, researchers conducting epithelial-bacterial in vitro co-culture should not underestimate
the effects of these parameters.
© 2017 Elsevier Inc. All rights reserved.