《Table 1 Inoculation of lactating cows with passage cultures of Mycoplasma leachii strain GN407 via

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《"Mycoplasma leachii causes bovine mastitis: Evidence from clinical symptoms,histopathology and immunohistochemistry"》

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1) CCU，color changing unit.2) IHC，immunohistochemistry.–，negative for detection using the corresponding assay；+，positive for detection using the corresponding assay，weakly positive；++，mildly or moderately positive；+++，severely or strongly positive.

For this study，seven lactating Holstein breed cows were selected.Before inoculation，milk was taken from all cows to exclude the presence of M.leachii by PCR（Chen et al.2017）；to exclude subclinical mastitis by somatic cell count，the presence of antibodies against M.leachii in serum was detected by indirect ELISA using the lipoprotein precursor A（LPPA）protein as the antigen（Wang et al.2014）.The cows selected met the following criteria:negative PCR detection of M.leachii in milk，somatic cell count in milk less than200 000 mL–1，and M.leachii-specific serum antibodies not detected by ELISA.Twelve quarters of six lactating cows（left fore and hind quarters of each cow）were inoculated with 5 mL of fresh culture containing 1×108 CCU mL–1 of M.leachii strain GN407 through intramammary ductal infusion，and another 12 quarters（right fore and hind quarters of each cow）were inoculated with heat-inactivated M.leachii culture as a control.As a negative control animal，two quarters of one lactating cow（left fore and hind quarters）were inoculated with 5 mL of heat-inactivated M.leachii culture，and another two quarters（right fore and hind quarters）were not inoculated with medium（Table 1）.Each infected cow and control cow were housed in separate pens，and they were fed and cared for in an accredited facility at the HVRI in accordance with uniform standard to prevent transmission between quarters and cows.