Differential Somatic Cell Count Research Articles

Genetic Parameters of Bovine Milk Fatty Acid Profile, Yield, Composition, Total and Differential Somatic Cell Count.

Simple SummaryIn the last years, consumers have shown a remarkable interest in the qualitative and healthy aspects of milk, leading to an increase of the demand for dairy products with high nutritional value. By applying a genetic approach, it is possible to improve the quality of the fat contained in milk and its derivatives. In the present study we explored the genetic aspects of bovine milk fatty acids, in order to evaluate the feasibility of including them in breeding programs to alter their proportions in milk and improve the nutritional and healthy quality of fat. Our results indicate that genetic selection on fatty acid profile is feasible, as fatty acids present sufficient genetic variability and are moderately heritable. However, in the light of estimated correlations with other milk traits, a clear breeding objective should first be established.The growing interest of consumers for milk and dairy products of high nutritional value has pushed researchers to evaluate the feasibility of including fatty acids (FA) in selection programs to modify milk fat profile and improve its nutritional quality. Therefore, the aim of this study was to estimate genetic parameters of FA profile predicted by mid-infrared spectroscopy, milk yield, composition, and total and differential somatic cell count. Edited data included 35,331 test-day records of 25,407 Italian Holstein cows from 652 herds. Variance components and heritability were estimated using single-trait repeatability animal models, whereas bivariate repeatability animal models were used to estimate genetic and phenotypic correlations between traits, including the fixed effects of stage of lactation, parity, and herd-test-date, and the random effects of additive genetic animal, cow permanent environment and the residual. Heritabilities and genetic correlations obtained in the present study reflected both the origins of FA (extracted from the blood or synthesized de novo by the mammary gland) and their grouping according to saturation or chain length. In addition, correlations among FA groups were in line with correlation among individual FA. Moderate negative genetic correlations between FA and milk yield and moderate to strong positive correlations with fat, protein, and casein percentages suggest that actual selection programs are currently affecting all FA groups, not only the desired ones (e.g., polyunsaturated FA). The absence of association with differential somatic cell count and the weak association with somatic cell score indicate that selection on FA profile would not affect selection on resistance to mastitis and vice versa. In conclusion, our findings suggest that genetic selection on FA content is feasible, as FA are variable and moderately heritable. Nevertheless, in the light of correlations with other milk traits estimated in this study, a clear breeding goal should first be established.

The Effect of Pre-milking Stimulation on Teat Morphological Parameters and Milk Traits in the Italian Water Buffalo.

Water buffaloes (Bubalus bubalis) are very sensitive to environmental stimulus before and during milking, and this explains why disrupted milk ejections due to blood oxytocin level instability are frequent in this species. According to the literature, the manual stimulation (MS) of teats before milking promotes oxytocin release and allows milk ejection to start within 2–3 min. However, the pre-milking stimulation of teats is not always part of the milking routine in Italian buffalo farms; moreover, buffaloes with unstable milk let-down are sometimes treated with exogenous oxytocin (OX). Different types of pre-milking stimulation can impact differently on the mammary gland epithelium and structures and, therefore, on milk yield. In this study, we observed the changes in teat morphological traits before and after the application of three types of stimuli, i.e., no manual stimulation (NS), MS, and OX, in buffaloes reared in an Italian dairy farm. In particular, measurements were available for 23 and 21 buffaloes for front and rear teats, respectively. Subsequently, the effect of the pre-milking stimulation type was estimated on teat morphological characteristics and on milk traits recorded after the application of stimuli. The results showed that the teat canal length was shorter (P < 0.05) after stimulation in the case of MS and OX compared to NS. Cistern diameter was overall greater for MS and lower for OX. On the contrary, teat wall thickness was greater and lower for OX and MS, respectively. Milk yield and quality (fat, protein, and somatic cell score) were similar across the three types of pre-milking stimulation. In perspective, the impact of these types of pre-milking stimulation should be evaluated on a large scale, and the focus might be put on mammary gland epithelium integrity, mastitis incidence, and other udder health indicators in milk, e.g., electrical conductivity, differential somatic cell count, lactose content, and sodium and chloride concentration.

Combining total and differential somatic cell count to screen for mastitis

Combining total and differential somatic cell count to screen for mastitis

Milk microbiome: evaluation study on the differences among cows with a different health status classified by leukocyte pattern

Milk microbiome: evaluation study on the differences among cows with a different health status classified by leukocyte pattern

Investigation of dairy cow performance in different udder health groups defined based on a combination of somatic cell count and differential somatic cell count

Mastitis is still the costliest disease in milk production. In particular, its subclinical form, which may spread unnoticeably within a herd, is a major challenge. Somatic cell count (SCC) is broadly used as an indicator for mastitis and thus the basis for udder health management programmes, e.g. through dairy herd improvement (DHI) testing. Since recently, differential somatic cell count (DSCC, representing the combined proportion of polymorphonuclear neutrophils and lymphocytes as a percentage of total SCC) is available in addition. Our study was aimed to investigate dairy cow performance in four newly defined udder health groups (UHG) based on SCC and DSCC results from DHI testing. In total, 961,835 test-day results generated in Austria, China, Estonia, Germany, and Spain between January 2019 and March 2020 were available for data analyses. Cows were categorised into four UHG depending on test day SCC and DSCC results (UHG A: healthy/normal, ≤200,000 cells/mL and ≤65 %; B: suspicious, ≤200,000 cells/mL and >65 %; C: mastitis, >200,000 cells/mL and >65 %; D: chronic/persistent mastitis, >200,000 cells/mL and ≤65 %). Linear mixed effect models were used to compare the performance of cows between the UHG based on the parameters milk weight, energy-corrected milk, fat, protein, lactose, and estimated milk value. Highest performance was found for cows in UHG A and ranged between 21.4 (Austria) and 38.3 kg per cow and day (Spain). Interestingly, cows in group B were significantly less productive (0.9–2.4% less daily milk production) compared to those in group A. Cows in groups C (6.0–9.8% less daily production compared to group A) and D were, as expected, even less productive with a particularly significant drop for cows in group D (17.5–38.5% less daily production). These trends could be observed in all countries involved in this study. Proportions of cows in the four different UHG differed between countries, changed slightly within countries depending on season, differed depending on parity and days in milk, and were seen to vary hugely between herds. In conclusion, this study demonstrates changes in performance of dairy cows depending on their udder health status as defined based on the combination of SCC and DSCC. In particular cows in UHG B and D are of interest as they cannot be identified working with SCC only. Nevertheless, the actual udder health management measures that could be taken based on the new UHG still require further investigation.

A simulation study to investigate the added value in using differential somatic cell count as an additional indicator for udder health management in dairy herds

Mastitis is one of the most costly diseases in dairy herds worldwide. Somatic cell count (SCC) is widely used as an indicator for subclinical intramammary infections (IMI) that may eventually cause mastitis in dairy herds. Differential somatic cell count (DSCC) has recently been introduced as an additional indicator for IMI. The objective of this study was to investigate the value of using DSCC as an additional indicator to select cows for testing and subsequent intervention for subclinical mastitis during the lactation. We parameterized an existing bio-economic simulation model for dairy herds to include DSCC. Then, we simulated three Danish dairy cattle herd situations with different pathogen distributions where the main pathogens were 1) Staphylococcus aureus, 2) Streptococcus agalactiae, and 3) Streptococcus uberis. In these herds, we simulated two different selection strategies for testing (bacterial culture) for subclinical IMI and various intervention strategies for test positive cases. The first selection strategy considered only SCC; cows were selected for testing if they had a low SCC measurement followed by two high SCC measurements. In the second selection strategy, cows additionally had to have a high DSCC measurement. Results showed that both selection strategies led to a similar net income and to a similar number of clinical and subclinical cases for all investigated intervention strategies. However, when using DSCC in the selection of animals, the number of treatment days and the number of cows culled in relation to IMI was reduced: The median annual number of treatment days was reduced by 25–38 days in herd 1, by 25–42 days in herd 2, and by 30–48 days in herd 3, depending on the intervention strategy. The median annual number of cows culled in relation to IMI was reduced by up to 8 cows (10 cows in herd 3) for one of the intervention strategies. Subject to limitations associated with model assumptions, these results suggest that considering DSCC when selecting cows for testing can reduce IMI related culling and the use of antibiotics without changing in-herd prevalence nor resulting in economic loss.

Evaluation of the new differential somatic cell count parameter as a rapid and inexpensive supplementary tool for udder health management through regular milk recording

Mastitis, particularly in its subclinical form, is the costliest disease in milk production causing substantial financial losses to the dairy industry, impairing animal welfare, and one of the main reasons for treating dairy cows with antimicrobials. Somatic cell count (SCC) is broadly used as an indicator for mastitis or intramammary infection (IMI) and is the basis for udder health management programmes, e.g., through monthly dairy herd improvement (DHI) testing. While SCC shows the total number of cells in milk, the new Differential SCC (DSCC) shows also the combined proportion of polymorphonuclear neutrophils (PMN) and lymphocytes as a percentage of the total SCC. In this study, we investigated the test characteristics of DSCC as a new supplementary indicator for mastitis screening. We collaborated with 11 herds totalling 969 dairy cows and collected metered DHI samples once a month over four months. The IMI status was assessed through analysis of aseptic composite hand-stripped samples using culture and followed by species identification using MALDI-ToF. The pathogens detected were categorised as ‘no’, ‘minor’, ‘major’, or ‘other’ pathogens. The results of our study showed that the DSCC parameter was significantly associated with the IMI status and the cow’s parity but not with days in milk or test-day milk weight. On the other hand, SCC was associated with all these four factors. DSCC counts were significantly higher in samples of cows with IMI caused by major pathogens as compared to cows with no IMI or IMI by minor or other pathogens. SCC alone, DSCC alone, and the combination of DSCC and SCC were further compared based on test characteristics using exemplary cut-offs. For example, working with a cut-off of 200,000 cells/ for SCC alone compared to working with the combination of DSCC of 65 % and/or 200,000 cells/mL to classify cows as infected by major pathogens, the sensitivity increased from 78 % to 92 % and the specificity decreased from 87 % to 66 %. With the combination, the positive predictive value changed from 52 % to 34 %, and the negative predictive value stayed at the same level (96 % vs 98 %). In summary, our study provides first insights on test characteristics of the DSCC parameter used in combination with the well-established SCC for monitoring udder health using DHI testing. This combination opens up the possibility to further improve udder health monitoring programmes (e.g., improved identification of IMI caused by major pathogens) but more work on the subject is needed.

Antibiotic dry cow therapy, somatic cell count, and milk production: Retrospective analysis of the associations in dairy herd recording data using multilevel growth models

Antibiotic dry cow therapy (DCT) is an important part of most mastitis control programs. Updating DCT recommendations is an ongoing topic due to the global problem of antimicrobial resistance. Finland, along with other Nordic countries, has implemented selective DCT for decades. Our study analyzed Dairy Herd Improvement (DHI) information from 241 Finnish farmers who participated in a survey about their drying-off practices. The aim was to evaluate herd-level associations between milk somatic cell count (SCC), milk production, and various antimicrobial DCT approaches both cross-sectionally in 2016 and longitudinally in 2012−2016. The three DCT approaches in the study were selective, blanket, and no DCT use. An additional aim was to evaluate whether dynamic changes occurred in herd-average SCC and annual milk production over five years, and whether these potential changes differed between different DCT approaches. The method for the longitudinal analyses was growth modeling with random coefficient models. Differences in SCC and milk production between farms with different DCT approaches were minor. Regardless of the farm’s DCT approach, annual milk production increased over the years, while average SCC was reasonably constant. The variability in SCC and milk production across all DCT groups was low between years, and most of the variability was between farms. Compared to other milking systems, farms with automatic milking system (AMS) had higher SCC, and in 2016 higher milk production. The results of this study suggest that it is possible to maintain low herd-average SCC and good milk production when using selective DCT and following the guidelines for prudent antimicrobial use. Average SCC and milk production varied across the herds, suggesting that advice on DCT practices should be herd-specific. The methodology of growth modeling using random coefficient models was applicable in analyzing longitudinal data, in which the time frame was relatively short and the number of herds was limited.

Differential Somatic Cell Count as a Novel Indicator of Milk Quality in Dairy Cows.

Simple SummaryRecently, high-throughput instruments have been used to analyze milk differential somatic cell count, represented by the combined proportion of polymorphonuclear leukocytes and lymphocytes, providing indirect information on the udder inflammation status of dairy cows. No information is available about the relationship between differential somatic cell count and milk quality, so the aim of this study was to investigate the effect of differential somatic cell count on the composition of a large number of individual milk samples. Results showed that milk quality worsened when differential somatic cell count was high. In particular, it was evidenced lower milk fat, protein, casein contents and casein index, and augmented milk fatty acids could be found with an increasing differential somatic cell count level. These findings confirmed that differential somatic cell count could be a new informative tool for dairy farmers to monitor the quality of milk.Recent available instruments allow to record the number of differential somatic cell count (DSCC), representing the combined proportion of polymorphonuclear leukocytes and lymphocytes, on a large number of milk samples. Milk DSCC provides indirect information on the udder health status of dairy cows. However, literature is limited regarding the effect of DSCC on milk composition at the individual cow level, as well as its relation to the somatic cell score (SCS). Hence, the aims of this study were to (i) investigate the effect of different levels of DSCC on milk composition (fat, protein, casein, casein index, and lactose) and (ii) explore the combined effect of DSCC and SCS on these traits. Statistical models included the fixed effects of days in milk, parity, SCS, DSCC and the interaction between SCS × DSCC, and the random effects of herd, animal within parity, and repeated measurements within cow. Results evidenced a decrease of milk fat and an increase in milk fatty acids at increasing DSCC levels, while protein, casein and their proportion showed their lowest values at the highest DSCC. A positive association was found between DSCC and lactose. The interaction between SCS and DSCC was important for lactose and casein index, as they varied differently upon high and low SCS and according to DSCC levels.

Randomized equivalence study comparing the efficacy of 2 commercial internal teat sealants in dairy cows

The use of an internal teat sealant (ITS) at dry-off has been repeatedly shown to improve udder health in the subsequent lactation. However, almost all ITS research conducted in North America has evaluated one product (Orbeseal, Zoetis, Parsippany, NJ). The objective of this study was to evaluate a new ITS product (Lockout, Boehringer-Ingelheim Animal Health, Duluth, GA), by comparing it directly to Orbeseal in a multi-site, randomized, positively controlled equivalence trial for health indicators during the dry period [quarter-level new intramammary infection (IMI) risk, IMI cure risk, and IMI risk at 1 to 13 d in milk, DIM] and during the first 100 DIM [clinical mastitis and culling or death risk and test-day milk somatic cell count (SCC) and milk yield]. At dry-off, cows were randomly allocated to be treated with Orbeseal or Lockout after blanket administration of a cloxacillin dry cow therapy product. Cows were then followed from dry-off until 100 DIM. Intramammary infection status at enrollment and at 1 to 13 DIM was determined using standard bacteriological methods, allowing for the measurement of IMI dynamics during the dry period (i.e., IMI cures and new IMI). The effect of ITS group on dry period IMI cure, dry period new IMI, and IMI risk at 1 to 13 DIM was determined using generalized linear mixed models (logistic). Marginal standardization was used to derive risk difference estimates. An equivalence hypothesis test was conducted to compare ITS groups for dry period new IMI risk (margin of equivalence was ±5% units). The effect of ITS group on clinical mastitis and culling or death was determined using Cox proportional hazards regression. The effect of ITS group on test-day SCC and milk yield was determined using linear mixed models. Final models indicated that measures of quarter-level IMI dynamics were similar between ITS groups (i.e., risk difference estimates and 95% confidence intervals all close to zero). Furthermore, Lockout was found to be equivalent to Orbeseal for dry period new IMI risk using an equivalence hypothesis test. Hazard ratio estimates for clinical mastitis and culling or death were close to 1 and differences in SCC and milk yield between ITS groups were close to 0, indicating negligible effects of ITS group on test-day SCC and milk yield. In most cases, these effect estimates were relatively precise (i.e., narrow 95% confidence intervals). We conclude that producers using blanket dry cow therapy could consider including Orbeseal or Lockout treatment in their programs.

Differential Somatic Cell Count as a Marker for Changes of Milk Composition in Cows with Very Low Somatic Cell Count.

Simple SummaryRecently, the availability of a high-throughput milk analyzer performing a partial differential somatic cell count (DSCC) opened new opportunities in investigations on bovine udder health. The information supplied by this new tool would be of importance in cows with a very low somatic cell count (SCC ≤ 50,000 cells/mL). Our investigation confirmed that the repeatability of the measurement allows its use under field conditions. The observational data did not find an association between DSCC and intramammary infections in very low SCC cows. However, our data showed that all the major milk components decreased as the DSCC was raised. These findings confirmed that DSCC could be a new informative tool for dairy farmers to monitor the changes in milk quality. DSCC may be suggested as a marker to identify early changes in milk composition, as a result of an alteration in milk secretion mechanisms.The recent availability of a high-throughput milk analyzer performing a partial differential somatic cell count (DSCC) opened new opportunities in investigations on bovine udder health. This analyzer has a potential limitation on the accuracy of measurements when the somatic cell count (SCC) is below 50,000 cells/mL, values characterizing a good proportion of lactating cows in many herds. We obtained data for cows below this threshold, assessed the repeatability of these measurements and investigated the relationship between DSCC and udder health, milk composition and yield. Overall, 3022 cow milk test records performed on a Fossomatic™ 7/DC (Foss A/S, Hillerød, Denmark) were considered; 901 of them had an SCC ≤ 50,000 cells/mL. These latter samples were analyzed by qPCR to identify the presence of bacteria. Overall, 20.75% of the samples (187) were positive. However, the health status did not have any significant association with DSCC. The analysis of the association of DSCC on milk fat, protein and casein showed a significant decrease in their proportions as the DSCC increased, whereas it was not observed for milk yield and lactose. Therefore, DSCC in very low SCC cows may be suggested as a marker to identify early changes in milk composition.

Antimicrobial Effects of Conditioned Medium From Amniotic Progenitor Cells in vitro and in vivo: Toward Tissue Regenerative Therapies for Bovine Mastitis.

There is increasing evidence to suggest that, in addition to their regenerative effect, mesenchymal stromal cells (MSCs), and their secretome have an anti-inflammatory and antimicrobial role in the innate immune response in conditions such as sepsis. However, there is no published information on the effect of MSCs in bovine mastitis. Mastitis often results in extensive tissue damage due to multi-microorganism co-infection. This study investigated the ability of amniotic-derived conditioned medium (CM), in vitro and in vivo, to counteract microbial action and restore healthy tissue capable of milk production. Following determination of a dose–response curve, 10,000 colony-forming units (CFU) of Staphylococcus aureus (S. aureus) were inoculated into bovine mammary epithelial cell culture with and without 10% CM (supplemented either at the time of bacteria inoculation or after 4 h). Acridine orange staining was used to assess cell viability/apoptosis. Additionally, an in vivo study was performed using 48 dairy cows with acute and chronic mastitis, treated with CM (treated group) or antibiotics (control group). In vitro results showed that CM can attenuate bacterial growth, as evaluated by the number of CFU. After 24 h of culture with S. aureus, 89.67% of mammary epithelial cells treated with CM were still alive, whereas all cells cultured without CM were dead. Rates of epithelial cell survival (60.67%) were similar when CM was added 4 h after bacteria inoculation. There was no difference in somatic cell count between cases of acute mastitis in the CM-treated or control group in the in vivo study. However, relapses in chronic mastitis were less common in the group receiving CM. Our results show that CM is able to mitigate bacterial growth in vitro and may be particularly useful in the treatment of chronic mastitis, aiding restoration of milk production in cows that would otherwise be removed from the production cycle.

Differential somatic cell count as an additional indicator for intramammary infections in dairy cows

Mastitis, often caused by intramammary infection (IMI), is a significant problem in dairy farming globally. Somatic cell count (SCC) is widely used as a parameter for screening IMI in cows that are then treated or culled. We investigated the potential of a new parameter, differential SCC (DSCC), to detect IMI at cow level when SCC is already known. We achieved this using bacterial culture (BC) and PCR to detect 4 categories of pathogens (major, minor, other, and any) in 2 Danish dairy herds. Quarter milk samples were collected from monthly dairy herd improvement samplings over 1 yr and analyzed with BC, whereas cow-level dairy herd improvement samples were analyzed using PCR. Days in milk, parity, and IMI status had a significant effect on DSCC. Using DSCC in addition to SCC significantly improved the indication of IMI compared with using only SCC in the any pathogen category in both herds as well as the minor pathogens category in herd 2 when BC was used for detection. When PCR was used to detect IMI, the use of DSCC in addition to SCC was significant for the other pathogens category in herd 1 and the minor pathogens category in herd 2. Thus, our data revealed that DSCC can add significant information describing IMI status even when SCC is already known; however, this depends on the causative pathogen. Future studies may address how to use DSCC in practice as well as consider the availability of temporal data to potentially gain insight into the course of infection.

Investigation of differential somatic cell count as a potential new supplementary indicator to somatic cell count for identification of intramammary infection in dairy cows at the end of the lactation period

The objective of this study was to investigate the new differential somatic cell count (DSCC) as a supplementary indicator to SCC for the identification of intramammary infection (IMI) in dairy cows at the end of the lactation period. Different approaches for identification of cows with IMI (i.e. often based on SCC) and targeted antimicrobial treatment of those rather than of all cows have been developed (i.e. selective dry cow treatment). Recently, DSCC representing the proportion of polymorphonuclear neutrophils and lymphocytes, has been introduced as an additional indicator for the presence of IMI. We used the last dairy herd improvement (DHI) samples taken within 42 d prior to dry-off as well as hand-stripped samples collected within 5 days prior to dry-off to measure DSCC and SCC. The bacteriological status was determined using quarter foremilk samples collected close to drying off. In total, 582 cows were dried off during our study but not all of them could be included in the data analysis for different reasons (e.g. incomplete data, samples too old for reliable determination of SCC and DSCC, contamination). Eventually, the final data set comprised of 310 cows of which 64 and 149 were infected with major and minor pathogens, respectively, and 97 were uninfected. The area under receiver-operating characteristics curves (AUC) were calculated to compare the diagnostic abilities of the different parameters. The AUC for identification of IMI by major pathogens when using the combination of DSCC and SCC was 0.64 compared to 0.62 for SCC alone and 0.62 for DSCC alone. The different parameters were further compared based on test characteristics and predictive values. For example, classifying cows as infected based on a cut-off of 200,000 cells/ml for SCC alone and in terms of using DSCC combined with SCC based on either >60% and/or >200,000 cells/ml, the sensitivity changed from 47 to 66% and the specificity from 74 to 54%. At the same time, the negative predictive value changed from 84 to 86% and the positive predictive value from 32 to 27%. Test characteristics and predictive values of the parameters DSCC and SCC were similar using DHI and hand-stripped samples. In conclusion, our study provides first indications on test characteristics and predictive values for the combination of DSCC and SCC. However, more work on this subject and the actual practical application is needed.

Short communication: Genetic aspects of milk differential somatic cell count in Holstein cows: A preliminary analysis

The aim of the present study was to assess genetic variation and heritability of a novel indicator of udder health, milk differential somatic cell count (DSCC), which represents the percentage of neutrophils plus lymphocytes in the total somatic cell count (SCC). Furthermore, we estimated genetic and phenotypic correlations of DSCC with other milk traits routinely measured in Italian Holstein cows. Besides DSCC, test-day data included milk yield, composition traits (i.e., fat, protein, casein, and lactose percentages), pH, milk urea nitrogen, and SCC. After editing, the final data set included 10,709 test-day records of 5,142 cows in 299 herds. Mean of DSCC was 62.07%, which means that macrophages were approximately 38% of total SCC. Comparing our results with the literature offered compelling evidence of the importance of acquiring information about the proportion of the different cell types in milk to better define the udder health status. In addition, our analysis revealed, for the first time, that DSCC is a heritable trait, and heritability (0.08 ± 0.02) was higher than that of traditional somatic cell score (0.04 ± 0.02). Nevertheless, heritability of DSCC is still low compared with milk yield and quality traits. Single-trait analysis within parity showed that DSCC was less heritable in primiparous than in multiparous cows, whereas bivariate analysis confirmed that DSCC and somatic cell score were 2 different traits, as their genetic and phenotypic correlations differed from unity. From a genetic point of view, the DSCC was positively weakly associated with milk yield, lactose percentage, and milk urea nitrogen, and negatively associated with pH. Our findings contributed to the understanding of the genetic background of DSCC and are a precursor to the potential use of DSCC in breeding programs to enhance cow resistance to mastitis. However, further research is needed to determine the weight this novel trait should receive in a selection program aimed to reduce udder health problems.

Milk cellularity and intramammary infections in primiparous and multiparous Lacaune ewes during early lactation

Despite the vast literature on postpartum mastitis in cattle, there is a demand for and lack of studies on the cellularity dynamics and cellular populations in milk and the occurrence of mastitis in ewes during the postpartum period. Thus, the aim of the present study was to evaluate the dynamics of cellularity in milk/colostrum and of mastitis in primiparous and multiparous Lacaune ewes during early lactation. For this purpose, 492 milk/colostrum samples were collected from the udder halves of 20 primiparous ewes and 21 multiparous ewes at the following six time points: day of parturition (T1) and one day (T2), three days (T3), seven days (T4), 15 days (T5), and 30 days (T6) after parturition. The milk and colostrum samples were subjected to microbiological examination, microscopic and automatic somatic cell counts (SCCs), differential somatic cell counts, and the California Mastitis test (CMT). The multiparous ewes had a higher prevalence of intramammary infections than the primiparous ewes during the postpartum period. Non-aureus staphylococci bacteria were the main mastitis etiological agents isolated in the present study. Furthermore, one of the most important findings of our study was that 35% of the udder halves of the primiparous ewes were infected at parturition, suggesting potential concerns for mastitis control programs. Finally, the SCC did not appear to be a good indicator of inflammation during early lactation because this measure was influenced by the parity and the sampling period. Moreover, the predictive values of the CMT and neutrophil counts were slightly better than those of the SCC during the immediate postpartum period.

Differential somatic cell count in milk before, during, and after lipopolysaccharide- and lipoteichoic-acid-induced mastitis in dairy cows

Intramammary infections induce the initiation of the inflammatory response, resulting in an increase in somatic cell count (SCC) in milk. The SCC includes several different types of cells but does not differentiate between them. On the contrary, the new differential somatic cell count (DSCC) parameter allows for the differentiation between 2 groups of cells: polymorphonuclear neutrophils (PMN) and lymphocytes versus macrophages. Therefore, the aim of this paper was to describe the changes of both DSCC and SCC during mastitis induced by cell wall components from typical mastitis-causing pathogens [lipopolysaccharide (LPS), Escherichia coli; lipoteichoic acid (LTA), Staphylococcus aureus] known to trigger different severities of mastitis. In addition, the effect the glucocorticoid prednisolone (PRED), which is known to attenuate the immune response in the mammary gland, was investigated. Twenty dairy cows were equally divided into 5 groups and treated with LPS, LTA, LPS+PRED, LTA+PRED, or a saline control. Milk samples were taken at the following time points: baseline (d -3, -2, and -1), right before treatment (d 0), 5 h after treatment (d 0.2), early cure phase (d 1 and 2), and late cure phase (d 3, 4, 5, 6, 7, and 14) and analyzed for DSCC and SCC. Mean DSCC values increased significantly from <60% at baseline and right before treatment to >81% 5 h after treatment and the early cure phase in all groups, except for the groups control and LTA+PRED. This increase clearly reflects a shift in cell populations to predominantly PMN. The SCC increased significantly following the stimulation, too, as expected. Interestingly, we observed cases where SCC increased moderately only whereas DSCC showed an evident increase, meaning that the shift in cell populations occurred even at low SCC levels. The PRED clearly lowered the cell migration in group LTA+PRED. This is the first ever study investigating DSCC during induced mastitis under controlled conditions. The combination of DSCC and SCC could be employed for the earlier detection of mastitis by revealing the shift in cell population independent from the SCC level. Furthermore, combining DSCC and SCC information could help to determine the stage of mastitis because we observed high DSCC and SCC results in the early stage of mastitis but evidently lower DSCC and high SCC in the cure phase. Hence, our results offer the first fundamental insights on how mastitis monitoring could be improved in the frame of dairy herd improvement programs.

Efficacy and clinical safety of pegbovigrastim for preventing naturally occurring clinical mastitis in periparturient primiparous and multiparous cows on US commercial dairies.

Periparturient dairy cows experience impaired immune function, exhibited as a transient decrease in neutrophil function. This decrease in immune competence is associated with an increase in susceptibility to bacterial infections, including mastitis and metritis. Bovine granulocyte colony stimulating factor (bG-CSF) is an endogenous protein that enhances neutrophil bactericidal functions and increases the production of neutrophils from bone marrow precursors. Administration of pegbovigrastim (recombinant bG-CSF covalently bound to polyethylene glycol) around the time of calving has been shown to reduce the incidence of new clinical mastitis cases in a natural disease model system. To further explore the application of pegbovigrastim under herd management systems typical of those found in the US dairy industry, we conducted a multicenter field study to evaluate the efficacy and clinical safety of pegbovigrastim administered to multiparous cows and heifers approximately 7 d before calving and within 24 h of calving. Responses of treated cows were compared with those of animals treated with sterile saline. Animals treated with pegbovigrastim exhibited 4- to 5-fold increases in circulating neutrophil numbers within 24 h of treatment initiation, and this increase persisted at least a week beyond the second dose. Pegbovigrastim-treated animals exhibited a 35% decrease in the incidence of clinical mastitis relative to the controls during the first 30 d of lactation. Animals treated with pegbovigrastim also exhibited a 52% reduction in failure to return to visual estrus within 80 d of calving. We observed no differences in somatic cell count or milk composition between treated and control animals. We also found no differences in the duration of pregnancy or proportion of viable calves in treated cows relative to control animals. These results indicate that administration of pegbovigrastim provides a well-tolerated, novel approach to overcoming periparturient immune suppression, resulting in reduced susceptibility to clinical mastitis during early lactation.

Differential somatic cell count—A novel method for routine mastitis screening in the frame of Dairy Herd Improvement testing programs

Somatic cell count (SCC) is used as key indicator in mastitis screening programs typically applied in the frame of Dairy Herd Improvement (DHI) testing programs. However, mastitis is still causing tremendous economic losses to the dairy industry. Hence, new biomarkers are needed that can be used for enhanced management of mastitis on dairy farms. Besides the determination of SCC, differentiation of cells has been described to be beneficial for a more definite description of the actual udder health status of dairy cows. The aim of this study was to develop a new method for rapid and simultaneous determination of SCC and a new parameter, differential somatic cell count (DSCC), in individual cow milk samples using flow cytometry. The method is sought to be applied in central milk testing laboratories, so that existing DHI infrastructures can be used. The DSCC represents the combined proportion of polymorphonuclear leukocytes (PMN) and lymphocytes expressed in percentage. The proportion of macrophages can be calculated by subtracting DSCC from 100%. Our research revealed increasing proportions of PMN, but decreasing proportions of macrophages as SCC increased. However, lymphocytes occurred fairly constantly with low proportions across the entire SCC range. Hence, the DSCC parameter reflects the antidromic trend of PMN and macrophages. Fluorescence microscopy was used to evaluate the specificity of the new Foss DSCC method in terms of DSCC and a high correlation was found. Apart from that, the accuracy of cell differentiation using the Foss DSCC method was confirmed in a cell sorting trial. Total SCC could be determined equally well using the new method as compared with existing methods. The new method was further proven to be robust toward a range of method and milk-sample-related factors. In an initial field trial, regular DHI samples of a local dairy herd were analyzed. The DSCC values occurred in a broad range from 34 to 79% in samples with <400,000 cells/mL. Higher DSCC values (53-89%) were found in samples with >400,000 cells/mL. In conclusion, the new Foss DSCC method allows reliable, repeatable, fast, robust, and accurate determination of both DSCC and SCC at low cost. This, in turn, provides more accurate information on the actual udder health status of dairy cows. The practical application of DSCC in the frame of DHI testing programs, however, needs further investigation.

Supraphysiological oxytocin increases the transfer of immunoglobulins and other blood components to milk during lipopolysaccharide- and lipoteichoic acid-induced mastitis in dairy cows.

Bacterial mastitis causes pathogen-dependent changes of the blood-milk barrier, and these changes can influence the differential transfer of blood components to milk. It is well known that gram-negative pathogens such as Escherichia coli can cause a greater activation of the immune system and thus a more comprehensive transfer of blood components including IgG than gram-positive pathogens such as Staphylococcus aureus. Supraphysiological doses of oxytocin (OT) have been shown to increase the permeability of the blood-milk barrier; however, the effect of OT during experimentally induced mastitis has not been investigated. Therefore, the objective of this study was to examine if intravenous administration of OT during lipopolysaccharide (LPS)- or lipoteichoic acid (LTA)-induced mastitis could influence the transfer of blood components to milk. The hypothesis was that OT could induce a greater transfer of blood components during mastitis. Twenty-seven dairy cows were injected via the teat canal with LPS, LTA, or a saline control followed by an intravenous injection of OT 2h following intramammary challenge. Milk samples were collected every half hour and analyzed for somatic cell count (SCC), IgG, lactate dehydrogenase (LDH), and serum albumin (SA). Due to the chosen dosage of LPS and LTA, there was no difference in SCC between quarters challenged with only LPS or LTA. Quarters challenged with LPS and OT had a higher SCC and a greater transfer of IgG, LDH, and SA compared with quarters challenged with only LPS. Quarters challenged with LTA and OT had a greater transfer of IgG, LDH, and SA, whereas the SCC increase did not differ from quarters only treated with LTA. In quarters treated only with OT, SCC, LDH, and SA increased, but no difference was observed in IgG concentration from untreated control quarters. In conclusion, there are pathogen-specific changes in the blood-milk barrier and OT can induce a greater transfer of blood components to milk in both LPS- and LTA-induced mastitis. Oxytocin could have implications for use as a mastitis therapy, as there was an increased transfer of IgG into the milk.