Relative mRNA level of immune factors expressed by milk cells compared to β-actin expression
| Positive group | Negative group | |||
|---|---|---|---|---|
| interleukin-6 | before | 11.80 ± 1.17a | 16.33 ± 0.68a | * |
| after | 7.97 ± 1.12b | 11.98 ± 0.71b | * | |
| lysozyme | before | 5.27 ± 0.34 | 5.39 ± 0.18 | |
| after | 5.13 ± 0.25 | 5.31 ± 0.25 | ||
| lactotransferrin | before | 9.28 ± 1.74a | 11.32 ± 0.75a | |
| after | 7.60 ± 0.97b | 8.42 ± 0.61b | ||
| defensin-β3 | before | 3.86 ± 0.27 | 2.99 ± 0.21 | |
| after | 3.60 ± 0.37 | 3.78 ± 0.18 | ||
| cathelicidin | before | 6.27 ± 0.56a | 8.24 ± 0.42a | * |
| after | 4.61 ± 0.74b | 5.86 ± 0.39b | ||
| pentraxin-3 | before | 4.31 ± 0.24 | 4.97 ± 0.22 | |
| after | 3.84 ± 0.35 | 4.20 ± 0.20 | ||
| caspase-3 | before | 13.84 ± 1.23 | 15.22 ± 0.30a | |
| after | 12.86 ± 0.92 | 13.49 ± 0.66b | ||
| elastase | before | 10.07 ± 0.96 | 11.19 ± 0.33 | |
| after | 10.043 ± 0.45 | 10.71 ± 0.25 |
Primers used for real-time PCR
| Gene | Accession | Product | Primer | Sequence (5'-3') |
|---|---|---|---|---|
| β-actin | NM_173979.3 | 76 | Forward | CCCAGATCATGTTCGAGACC |
| Reverse | GAGGCATACAGGGACAGCAC | |||
| interleukin-6 | NM_173923.2 | 102 | Forward | AGATCCTGAAGCAAAAGATCGC |
| Reverse | TTGCGTTCTTTACCCACTCGT | |||
| lysozyme | NM_001078159.1 | 104 | Forward | TTCGGTCGCTGTCCAAGGCAAGGT |
| Reverse | TCCAGTTTGCCAGGCTGATTCCCC | |||
| Forward | CACCGAGCACTGGATAAAGGG | |||
| lactotransferrin | NM_180998.2 | 136 | Reverse | AGCCAGACACAGTCCAAGGG |
| Forward | GCATACAAGTGACTGCCCCTG | |||
| defensin-β3 | NM_001282581.1 | 70 | Reverse | ACTGAACCTCTTCCCTGTGTG |
| cathelicidin | Y09472.1 | 70 | Forward | GAAAGACTCTCCTACCCTGCCC |
| Reverse | CTCTAAGGGACACACAGCCAGG | |||
| pentraxin-3 | NM_001076259.2 | 80 | Forward | GCAGAGTCCTGTCACATCCG |
| Reverse | ACATACTGTGCTCCTCCGTG | |||
| caspase-3 | NM_001077840.1 | 148 | Forward | CAGGAGTGGAGTCCAGCAGA |
| Reverse | TCCAGAGTCCATTGATTTGCTTC | |||
| elastase | NM_001105653.1 | 119 | Forward | TTGCCTCTGTCGCCAAGTTT |
| Reverse | ACAGCCTTTCTAGTGGGTCC |
Major bacteria detected before and after milking in the positive group (n = 12)
| aerobic culture | anaerobic culture | |||
|---|---|---|---|---|
| before | after | before | after | |
| Aerococcus viridans | 3 | 1 | 2 | 1 |
| Staphylococcus chromogenes | 1 | 1 | 1 | 1 |
| Staphylococcus haemolyticus | 3 | 2 | 2 | 1 |
| Staphylococcus xylosus | 3 | 3 | 4 | 4 |
| Streptococcus dysgalactiae | 0 | 1 | 1 | 1 |