The main objective was to determine the prevalence of intramammary infections (IMI) in dairy cows in Flanders, Belgium. Data were obtained from quarter milk samples of dairy herds subjected to a mandatory yearly screening of all lactating cows. A total of 178 668 quarter milk samples were collected at 1087 cross-sectional dairy herd screenings performed in three consecutive years. Of the dairy cows, 40% had at least one culture-positive quarter. More than 50% of all IMI were caused by non-aureusstaphylococci.Streptococcus agalactiaeis almost eradicated in Flanders, whereasStaphylococcus aureuswas isolated from 18% of the culture-positive quarters. In addition, the distribution of mastitis pathogens in quarter milk samples from selected dairy cows with an elevated somatic cell count (SCC) is described. From 6390 cows with a geometric mean composite SCC ⩾250 000 cells/ml, nearly 65% had at least one culture-positive quarter. The majority of the IMI were caused by non-aureusstaphylococci (41·1%), whereasStaph. aureusand aesculin-positive cocci were found in respectively 25% and 18% of the culture-positive milk samples. We conclude that more efforts are needed in the prevention and control of subclinical mastitis in Flanders. Non-aureusstaphylococci are the predominant cause of IMI, warranting more research regarding the epidemiology and pathogenicity of those species.

Intramammary infections (IMI) in recently calved dairy heifers are more common than was formerly believed but their relevance for future performance has been studied only rarely. In the present study, the association between the IMI status of fresh heifers and their subsequent udder health, milk production, and culling in first lactation was explored. Quarter milk samples were collected between 1 and 4 d in milk (DIM) and between 5 and 8 DIM from 191 dairy heifers in 20 dairy herds for bacteriological culturing and somatic cell count (SCC) analysis. Monthly milk recording data including composite milk SCC and test-day milk yield (MY) were obtained for the first 285 DIM or until culling. Farmer-recorded clinical mastitis cases were available. Data were analyzed using mixed models and survival analysis. Approximately 80% of the fresh heifers (79.8%) had at least one culture-positive quarter. Coagulase-negative staphylococci (CNS) were the most frequently isolated pathogens (72%), followed by esculin-positive streptococci (4.6%) and Staphylococcus aureus (3.5%). Overall geometric mean SCC at quarter level decreased between the first and second samplings from 348,000 to 116,000 cells/mL. Heifers infected with CNS had an intermediate average test-day SCC (84,000 cells/mL) during the first 285 DIM compared with noninfected heifers (53,000 cells/mL) and heifers infected with major pathogens (195,000 cells/mL). Heifers infected with major pathogens had a much lower average daily MY (18.3kg) during first lactation compared with noninfected animals (21.3kg). That CNS-infected heifers out-produced their noninfected counterparts could be at least partially explained by their significantly lower incidence of clinical mastitis (incidence risk 3.6 vs. 21.0%) during first lactation compared with noninfected heifers. We conclude that although CNS cause the majority of IMI in heifers around calving, they should not be a reason for serious concern.

Introduction: Mesenchymal stromal cells (MSCs) have been extensively studied for their promising capabilities in regenerative medicine. Although bone marrow is the best-known source for isolating equine MSCs, non-invasive alternative sources such as umbilical cord blood (UCB), umbilical cord matrix (UCM), and peripheral blood (PB) have also been reported. Methods: Equine MSCs from three non-invasive alternative sources were isolated from six individual mares (PB) and their foals (UCB and UCM) at parturition. To minimize inter-horse variability, the samples from the three sources were matched within the same mare and for UCB and UCM even within the same foal from that specific mare. The following parameters were analyzed: (i) success rate of isolation, (ii) proliferation capacity, (iii) tri-lineage differentiation ability, (iv) immunophenotypical protein, and (v) immunomodulatory mRNA profiles. Linear regression models were fit to determine the association between the source of MSCs (UCB, UCM, PB) and (i) the moment of first observation, (ii) the moment of first passage, (iii) cell proliferation data, (iv) the expression of markers related to cell immunogenicity, and (v) the mRNA profile of immunomodulatory factors, except for hepatocyte growth factor (HGF) as no normal distribution could be obtained for the latter variable. To evaluate the association between the source of MSCs and the mRNA expression of HGF, the non-parametric Kruskal-Wallis test was performed instead. Results: While equine MSCs could be isolated from all the UCB and PB samples, isolation from UCM was successful in only two samples because of contamination issues. Proliferation data showed that equine MSCs from all three sources could be easily expanded, although UCB-derived MSCs appeared significantly faster in culture than PB- or UCM-derived MSCs. Equine MSCs from both UCB and PB could be differentiated toward the osteo-, chondro-, and adipogenic lineage, in contrast to UCM-derived MSCs in which only chondro- and adipogenic differentiation could be confirmed. Regardless of the source, equine MSCs expressed the immunomodulatory genes CD40, CD80, HGF, and transforming growth factor-beta (TGFβ). In contrast, no mRNA expression was found for CD86, indoleamine 2,3-dioxygenase (IDO), and tumor necrosis factor-alpha (TNFα). Conclusions: Whereas UCM seems less feasible because of the high contamination risks and low isolation success rates, UCB seems a promising alternative MSC source, especially when considering allogeneic MSC use.

The objective of this study was to further scrutinize the previously found positive association between intramammary infection (IMI) caused by coagulase-negative staphylococci (CNS) in early lactating heifers and test-day daily milk yield (MY) throughout first lactation, with a specific focus on the effect of the heifers' genetically determined milk production levels and the incidence of clinical mastitis. Two precise longitudinal data sets were analyzed using a series of statistical models including potential confounding and intermediate variables. The final database included the IMI status at calving, composite milk somatic cell count (SCC) and MY records at test day up to 285 d in milk (DIM), farmer-recorded clinical mastitis (CM) cases between 14 and 285 DIM, estimated new IMI incidence based on a SCC threshold of 100,000 cells/mL between 14 and 285 DIM, DIM, average 305-d MY at the herd level, and the heifers' genetic merit for MY from 240 dairy heifers from 29 dairy herds. Seventy-one (29.6%) early lactating heifers were noninfected, 108 heifers (45.0%) were CNS infected, and 61 heifers (25.4%) were infected with any major pathogen. The positive effect of CNS IMI in early lactation on test-day MY was estimated at 1.32 kg/d using a first basic mixed regression model. Correcting for the confounder genetic merit for MY reduced this effect to 1.17 kg. Interestingly, taking into account the confounding effect of herd resulted in an increase of the estimate from 1.32 to 2.2 kg/d. The positive effect of CNS IMI in early lactation on MY after correcting the model for both confounders was estimated at 2.05 kg/d. Heifers infected with CNS in the first DIM tended to have fewer CM cases throughout lactation compared with the noninfected herd mates. Including the intermediate variable CM in the model explained 0.16 kg/d of the corrected effect of 2.05 kg/d. Inclusion of test-day SCC, another intermediate variable, however, increased the estimate by 0.11 kg/d. With an appropriate correction for several confounders and biologically understood intermediate variables such as CM, test-day SCC, and new IMI based on SCC threshold of 100,000 cells/mL, an unexplained test-day MY difference between CNS-infected and noninfected heifers of 2.0 kg/d remained.