Subclinical mastitis is one of the major health problems in dairy herds due to decreased milk production and reduced milk quality. The aim of this study was to examine the within-herd prevalence of subclinical intramammary infection caused by Mycoplasma bovis and to evaluate associations between M. bovis and cow daily milk yield, udder health, and milk composition. Individual cow composite milk samples (n = 522) were collected from all lactating dairy cows in 1 Estonian dairy farm in November 2014. Daily milk yield, days in milk, and parity were recorded. Collected milk samples were analyzed for somatic cell count, milk protein, fat, and urea content. The presence of M. bovis, Staphylococcus aureus, Streptococcus agalactiae, and Streptococcus uberis in the milk samples was confirmed by quantitative PCR analysis. The within-herd prevalence of M. bovis was 17.2% in the study herd. No association was observed between days in milk and parity to the presence of M. bovis in milk. According to linear regression analysis, the daily milk yield from cows positive for M. bovis was on average 3.0 kg lower compared with cows negative for M. bovis. In addition, the presence of M. bovis in milk samples was significantly associated with higher somatic cell count and lower fat and urea content compared with milk samples negative for M. bovis. In conclusion, subclinical M. bovis intramammary infection is associated with decreased milk yield and lower milk quality.
We aimed to evaluate the elimination of 4 different mastitis pathogens, Streptococcus agalactiae, Mycoplasma bovis, Staphylococcus aureus, and Streptococcus uberis, from infected udder quarters during the dry period using quantitative PCR. The second purpose of this study was to evaluate the association between milk haptoglobin (Hp) concentration and the presence of udder pathogens (Strep. agalactiae, Staph. aureus, M. bovis, and Strep. uberis) in udder quarter milk samples before and after dry period. Aseptic udder quarter milk samples (n = 1,001) were collected from 133 dairy cows at dry off and at the first milking after calving from 1 large dairy herd. Bacterial DNA of Strep. agalactiae, Staph. aureus, Strep. uberis, and M. bovis in the udder quarter milk samples was identified with commercial quantitative PCR analysis Mastitis 4B (DNA Diagnostic A/S, Risskov, Denmark). Milk Hp concentration (mg/L) was measured from udder quarter milk samples. The elimination rates during the dry period for M. bovis, Staph. aureus, Strep. agalactiae, and Strep. uberis were 86.7, 93.6, 96.2, and 100.0%, respectively. The new IMI rate was 3.0% for M. bovis, 2.9% for Staph. aureus, 2.4% for Strep. agalactiae, and 3.1% for Strep. uberis. The milk Hp concentration was significantly higher in udder quarter milk samples with blood and in samples positive for Strep. agalactiae at dry off and for Staph. aureus postcalving. Elevated milk Hp concentration was not associated with the presence of M. bovis in the udder quarter milk samples. In conclusion, elimination of Staph. aureus, Strep. agalactiae, and Strep. uberis during the dry period was high; the elimination of M. bovis from infected udder quarters was lower, but probably spontaneous. Additionally, milk Hp concentration may be used as a marker for udder inflammation when combined with the bacteriological results at dry off and postpartum.
Milk quality in bulk tank milk (BTM) is measured by flow cytometry technology as total bacterial count (TBC) and somatic cell count (SCC). To investigate SCC problems, culture or PCR can be used to identify mastitis causing bacteria, e.g., Mastit 4, a commercially available qPCR test. TBC in BTM can be investigated further using culture-based methods such as standard plate count, laboratory pasteurization count, coliform count, and spore counts. To our knowledge, no qPCR addressing the bacteria involved in TBC has been commercially introduced. The aim of this study is to evaluate a recently introduced 3-h qPCR test, TBC 4. The TBC 4 qPCR detects four target groups, Pseudomonas, Streptococci, Enterobacteriacea/ Enterococcus, and Bacillus/Clostridia. These target groups relates to problems on the farm such as cooling, mastitis, environment, and silage. We will continue with new research to compare the TBC 4 qPCR test with traditional culture. For this study, BTM samples from different TBC intervals were selected based on BactoCount results found at routine payment investigation at Eurofins laboratory (Vejen, Denmark). These samples were analyzed using TBC 4 qPCR assay within 24 h. In total 346 BTM samples were divided into 6 different intervals of colony forming units (CFU). For all four targets in each of the different intervals of CFU, the percent of positive samples, the average C t-value, the percent of positive samples with C t <30 and C t <25 were calculated. For Pseudomonas, Streptococci, and Enterobacteriacea/Enterococcus the number of positive samples with lower C t-values (high bacteria content) correlated with the CFU mL-1. We found Enterobacteriacea/ Enterococcus, Pseudomonas, and Streptococci in high number of bacteria (C t <25) in 25, 19 and 56% of samples with CFU mL-1 between 50 001-100 000 and 53, 44, and 39% in samples with CFU mL-1 >100 000. The TBC 4 qPCR test showed to be a strong and fast tool for farmers, advisors and service technicians to address problems with high TBC and ensuring the delivery of good quality milk to the dairy.
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