Comparing effects of bovine Streptococcus and Escherichia coli mastitis on impaired reproductive performance.

In 2 epidemiological studies, we evaluated the effect of mastitis induced by gram-positive Streptococcus and gram-negative Escherichia coli on impaired reproductive performance in lactating Holstein cows. In the first study, 52,202 cows from 178 dairy farms throughout Israel were divided into groups based on infection before first artificial insemination (AI) with Streptococcus or E. coli, 3 groups with elevated somatic cell count (SCC) without infection by those pathogens [low SCC (200-400) × 103 cell/mL; medium SCC (401-1,000) × 103 cell/mL; high SCC, >1,000 × 103 cell/mL], and uninfected controls. Pregnancy per first AI (P/1stAI) and pregnancy rate at 300 d in milk (PREG 300) were analyzed by the GLIMMIX procedure (SAS); number of AI per pregnancy (AI/P), days open, and rest days (calving to first AI) were analyzed by the MIXED procedure (SAS Institute Inc., Cary, NC). Values of P/1stAI were similarly low for Streptococcus and E. coli (27-28%) versus 42% in controls; PREG 300 was lower for Streptococcus (76%) than for E. coli (79%) versus 88% for uninfected controls and a mean 83% for the elevated SCC groups. Days open and number of AI/P were higher than in controls and similar in Streptococcus and E. coli groups. The second study included 778 cows on 6 dairy farms; the cows were infected before first AI by Streptococcus or E. coli or uninfected. Resumption of cyclicity was determined by an automated activity-monitoring system, and data were sorted by time of infection before or after cyclicity resumed. The Streptococcus group had lower P/1stAI before and after cyclicity (26 and 27%, respectively) than the E. coli group (31 and 34%, respectively) and uninfected controls (42%). Notably, PREG 300 in the Streptococcus group before (73%) and after (67%) cyclicity was much lower than for the E. coli group (85 and 93%, respectively) and the controls (95%). A marked rise in day of cyclicity resumption (∼80 d) was observed in cows that were infected early on. Number of AI/P was higher in the mastitic groups than in uninfected controls. Uterine disease postpartum, although more prevalent among Streptococcus cows, did not substantially alter the larger reduction in P/1stAI and PREG 300 in Streptococcus versus E. coli cows. Thus, long-term Streptococcus-induced mastitis disrupted fertility more than short-term acute E. coli-induced mastitis, resulting in a much higher percentage of Streptococcus cows in late lactation that did not conceive due to reproduction failure.

Distribution of indications and procedures within the framework of centers participating in the WAA apheresis registry.

The WAA apheresis registry was established in 2003 and an increasing number of centers have since then included their experience and data of their procedures. The registry now contains data of more than 74,000 apheresis procedures in more than 10,000 patients. This report shows that the indications for apheresis procedures are changing towards more oncological diagnoses and stem cell collections from patients and donors and less therapeutic apheresis procedures. In centers that continue to register, the total extent of apheresis procedures and patients treated have expanded during the latest years.

Real-time evaluation of individual cow milk for higher cheese-milk quality with increased cheese yield.

Cheese was produced in a series of experiments from milk separated in real time during milking by using the Afilab MCS milk classification service (Afikim, Israel), which is installed on the milk line in every stall and sorts milk in real time into 2 target tanks: the A tank for cheese production (CM) and the B tank for fluid milk products (FM). The cheese milk was prepared in varying ratios ranging from ~10:90 to ~90:10 CM:FM by using this system. Cheese was made with corrected protein-to-fat ratio and without it, as well as from milk stored at 4°C for 1, 2, 3, 4, and 8d before production. Cheese weight at 24h increased along the separation cutoff level with no difference in moisture, and dry matter increased. The data compiled allowed a theoretical calculation of cheese yield and comparing it to the original van Slyke equation. Whenever the value of Afi-Cf, which is the optical measure of curd firmness obtained by the Afilab instrument, was used, a better predicted level of cheese yield was obtained. In addition, 27 bulk milk tanks with milk separated at a 50:50 CM:FM ratio resulted in cheese with a significantly higher fat and protein, dry matter, and weight at 24h. Moreover, solids incorporated from the milk into the cheese were significantly higher in cheeses made of milk from A tanks. The influence of storage of milk up to 8d before cheese making was tested. Gross milk composition did not change and no differences were found in cheese moisture, but dry matter and protein incorporated in the cheese dropped significantly along the storage time. These findings confirm that milk stored for several days before processing is prone to physico-chemical deterioration processes, which result in loss of milk constituents to the whey and therefore reduced product yield. The study demonstrates that introducing the unknown parameters for calculating the predicted cheese yield, such as the empiric measured Afi-Cf properties, are more accurate and the increase in cheese yield is more than increasing just the protein level, the value that is being tested by the dairies, or even casein.

Adverse events in apheresis: An update of the WAA registry data.

Apheresis with different procedures and devices are used for a variety of indications that may have different adverse events (AEs). The aim of this study was to clarify the extent and possible reasons of various side effects based on data from a multinational registry. The WAA-apheresis registry data focus on adverse events in a total of 50846 procedures in 7142 patients (42% women). AEs were graded as mild, moderate (need for medication), severe (interruption due to the AE) or death (due to AE). More AEs occurred during the first procedures versus subsequent (8.4 and 5.5%, respectively). AEs were mild in 2.4% (due to access 54%, device 7%, hypotension 15%, tingling 8%), moderate in 3% (tingling 58%, urticaria 15%, hypotension 10%, nausea 3%), and severe in 0.4% of procedures (syncope/hypotension 32%, urticaria 17%, chills/fever 8%, arrhythmia/asystole 4.5%, nausea/vomiting 4%). Hypotension was most common if albumin was used as the replacement fluid, and urticaria when plasma was used. Arrhythmia occurred to similar extents when using plasma or albumin as replacement. In 64% of procedures with bronchospasm, plasma was part of the replacement fluid used. Severe AEs are rare. Although most reactions are mild and moderate, several side effects may be critical for the patient. We present side effects in relation to the procedures and suggest that safety is increased by regular vital sign measurements, cardiac monitoring and by having emergency equipment nearby.

Two approaches to improve fertility of subclinical mastitic dairy cows.

Mastitis, particularly in its subclinical form, is a widely spread disease that reduces the fertility of lactating cows. A major cause of poor conception risk has been associated with delayed ovulation of a large subgroup of subclinical mastitic cows. This study examined 2 approaches to improve fertility in this subgroup. Subclinical mastitic cows were defined by somatic cell count elevated above a threshold of 150,000 cells/mL of milk determined in all monthly test day samples collected before AI. Uninfected (control) cows were defined by somatic cell count below threshold. In experiment 1, we examined a hormonal approach aimed to correct the timing of ovulation in mastitic cows in which it would otherwise be delayed. The probability of conception of mastitic and uninfected groups following Ovsynch (OVS) and timed AI versus AI following detected estrus (E) was examined (n=1,553 AI) and analyzed by a multivariable, logistic model statement using the GLIMMIX procedure of SAS. The OVS protocol significantly elevated the probability of conception of mastitic cows to a level similar to that of their uninfected counterparts. Actual mean conception risks for uninfected-E, subclinical-E, uninfected-OVS, and subclinical-OVS groups were 41.8, 26.4, 39.3, and 40.5%, respectively. The OVS protocol did not improve probability of conception in cows diagnosed with uterine disease postpartum. In experiment 2, a management approach aimed to better synchronize timing of ovulation with timing of AI in subclinical mastitic cows was examined. A second AI was added 24h after the first (routine) AI, following detection of natural estrus. Probability of conception did not differ between subclinical mastitic cows inseminated once or twice. Lack of improvement in conception risk might be related to low preovulatory LH surge in mastitic cows, which is likely to induce not only delayed ovulation but also disruption of oocyte maturation. Thus the OVS protocol can improve fertility of subclinical mastitic cows, probably due to "corrected" timing of ovulation in cows in which it would otherwise be delayed.

Additive solutions differentially affect metabolic and functional parameters of platelet concentrates.

BACKGROUND: Pathogen inactivation (PI) of platelet concentrates with extension of shelf life to 7 days requires the use of platelet additive solutions (PAS). We examined the quality of platelets resuspended in three different PAS stored for up to 7 days. MATERIALS AND METHODS: Twelve triple adult dose platelet concentrates (PC) were collected using the TrimaAccel® collection system. Each highly concentrated product was divided into three equal parts, and the additive solutions (Composol® or SSP+® or Intersol™) were added to a final concentration of 56% PAS and 44% plasma. Samples were drawn on days 1, 5 and 7 to measure pH, glucose, lactate dehydrogenase (LDH), lactate, mean platelet volume (MPV) and the aggregation response to collagen and the thrombin receptor agonist peptide-6. Further, p-selectin expression on platelets was assessed. RESULTS: No statistically significant changes were observed for pH and MPV during 7 days of storage in all PAS containing PCs, whereas glucose decreased and LDH and lactate increased over time (P < 0·05). These changes were particularly evident in Intersol PCs on days 5 and 7 compared with Composol® PCs or SSP+® PCs (P < 0·05). Platelets from Intersol PCs exhibited the highest baseline activation of p-selectin and showed reduced collagen- and TRAP-6-induced aggregation. CONCLUSION: Resuspension of platelets in Intersol for 7 days results in increased platelet activation and platelet metabolism compared with SSP+® or Composol®. Further clinical studies are needed to evaluate whether the observed differences in PAS-PCs affect the recovery rate or the life span of transfused platelets.

Subclinical mastitis disrupts oocyte cytoplasmic maturation in association with reduced developmental competence and impaired gene expression in preimplantation bovine embryos.

Subclinical chronic mastitis was induced to examine the effects on oocyte developmental competence. Uninfected Holstein cows were intramammary administrated with serial (every 48h for 20 days) low doses of toxin of Staphylococcus aureus origin (Gram-positive; G+), endotoxin of Escherichia coli origin (Gram-negative; G-) or sterile saline (control). Follicular fluid of toxin- and saline-treated cows was aspirated from preovulatory follicles and used as maturation medium. Oocytes harvested from ovaries collected at the abattoir were matured and then fertilised and cultured for 8 days. The percentage of oocytes undergoing nuclear maturation, determined by meiotic nuclear stages, did not differ between groups. Cytoplasmic maturation, determined by cortical granule distribution, was affected by both toxins (PPPPTGS2) mRNA increased, whereas that of growth differentiation factor 9 (GDF9) decreased in matured oocytes. In addition, PTGS2 expression increased and POU class 5 homeobox 1 (POU5F1) expression decreased in 4-cell embryos developed from both G+ and G- oocytes. Thus, regardless of toxin type, subclinical mastitis disrupts oocyte cytoplasmic maturation and alters gene expression in association with reduced developmental competence.

LGR5 rs17109924 is a predictive genetic biomarker for time to recurrence in patients with colon cancer treated with 5-fluorouracil-based adjuvant chemotherapy.

We recently found variants in cancer stem cell genes (CD44, ALCAM and LGR5) significantly associated with increased time to recurrence (TTR) in patients with stage III and high-risk stage II colon cancer treated with 5-fluorouracil (5-FU)-based chemotherapy. In this study, we validated these genetic biomarkers in a large and independent patient cohort (n=599). Patients who received 5-FU-based adjuvant chemotherapy (n=391) carrying at least one C allele in LGR5 rs17109924 had a significantly increased TTR compared with patients carrying the homozygous T/T variant (HR 0.38, 95%CI 0.19-0.79; P=0.006). In patients treated with surgery alone (n=208), no association between LGR rs17109924 and TTR was found (P=0.728). In the multivariate Cox-analysis, LGR5 rs17109924 remained statistically significant (HR 0.38, 95%CI 0.18-0.78; P=0.008) for patients who received adjuvant chemotherapy. We confirmed in a large and independent study cohort that LGR5 rs17109924 is a predictive genetic biomarker for TTR in patients with colon cancer treated with 5-FU-based adjuvant chemotherapy.

Host-specificity of Staphylococcus aureus causing intramammary infections in dairy animals assessed by genotyping and virulence genes.

Staphylococcus aureus is one of the most relevant pathogens causing clinical and subclinical, chronic mastitis in dairy animals. Routinely, mastitis pathogens are isolated and classified to genus or species level, and regarded as single entities. However, S. aureus includes a broad range of genotypes with distinct pathogenic and epidemiologic characteristics. The objective of the present study was to assess the host-specificity of S. aureus causing mastitis in dairy animals, based on phylogenetic and genotypic characterization as well as the presence of virulence and antimicrobial resistance genes in the pathogen genome. S. aureus isolates from mastitis in cows, sheep and goats in Israel, and from cows in Germany, the USA and Italy, were compared by the following methods: a. Bayesian phylogenetic comparison of sequences of genes nuc, coa, lukF and clfA, b. genotyping by spa and agr typing, and assignment to MLST Clonal Complexes (MLST CC), and c. the presence of a broad array of virulence and antimicrobial resistance genes. Overall, phylogenetic, virulence and genotyping approaches agreed with each other. Cow isolates could be differentiated from sheep and goat isolates with all three methods, with different resolution. In two phylogenetic clusters, segregation was found also between cow isolates from Israel and abroad. Sheep and goats' isolates showed less variability than isolates from cows in all methods used. In conclusion, different S. aureus lineages are associated to cows in contrast to goats and sheep, suggesting co-evolution between pathogen and host species. Modern diagnostics approaches should aim to explore molecular data for a better understanding and cost-effective management of mastitis.

Identifying the major bacteria causing intramammary infections in individual milk samples of sheep and goats using traditional bacteria culturing and real-time polymerase chain reaction.

Use of DNA-based methods, such as real-time PCR, has increased the sensitivity and shortened the time for bacterial identification, compared with traditional bacteriology; however, results should be interpreted carefully because a positive PCR result does not necessarily mean that an infection exists. One hundred eight lactating dairy ewes (56 Manchega and 52 Lacaune) and 24 Murciano-Granadina dairy goats were used for identifying the main bacteria causing intramammary infections (IMI) using traditional bacterial culturing and real-time PCR and their effects on milk performance. Udder-half milk samples were taken for bacterial culturing and somatic cell count (SCC) 3 times throughout lactation. Intramammary infections were assessed based on bacteria isolated in ≥2 samplings accompanied by increased SCC. Prevalence of subclinical IMI was 42.9% in Manchega and 50.0% in Lacaune ewes and 41.7% in goats, with the estimated milk yield loss being 13.1, 17.9, and 18.0%, respectively. According to bacteriology results, 87% of the identified single bacteria species (with more than 3 colonies/plate) or culture-negative growth were identical throughout samplings, which agreed 98.9% with the PCR results. Nevertheless, the study emphasized that 1 sampling may not be sufficient to determine IMI and, therefore, other inflammatory responses such as increased SCC should be monitored to identify true infections. Moreover, when PCR methodology is used, aseptic and precise milk sampling procedures are key for avoiding false-positive amplifications. In conclusion, both PCR and bacterial culture methods proved to have similar accuracy for identifying infective bacteria in sheep and goats. The final choice will depend on their response time and cost analysis, according to the requirements and farm management strategy.

Effects of Escherichia coli- and Staphylococcus aureus-induced mastitis in lactating cows on oocyte developmental competence.

Mastitis is associated with decreased fertility in dairy cows. In the current study, we created an experimental model to simulate short-term mastitis by a single intramammary administration of Gram-negative endotoxin of Escherichia coli origin (G-), or Gram-positive toxin of Staphylococcus aureus origin (G+), to examine the effect of mastitis on oocyte developmental competence. Healthy Holstein cows were synchronized, and follicular fluid (FF) of cows treated with G+ or G- and of uninfected cows (controls) was aspirated from the preovulatory follicles by transvaginal ultrasound procedure. The aspirated FF was used as maturation medium for in vitro embryo production. The distribution of matured oocytes into different cortical granule classes and meiotic stages was affected by G- administration (P<0.05) but not by G+ administration. The proportion of oocytes that cleaved to two- and four-cell stage embryos (44 h postfertilization) was lower in both G+ and G- groups than in controls (P<0.05). Blastocyst formation rate (7-8 days postfertilization) was lower in the G- group (P<0.05) and numerically lower in the G+ group compared with their uninfected counterparts. The total cell number in blastocysts did not differ among groups; however, the apoptotic index was higher in the G+ group (P<0.05), but not in the G- group, relative to controls. Examining mRNA relative abundance in oocytes and early embryos revealed mastitis-induced alterations in PTGS2 (COX2), POU5F1, and HSF1 but not in SLC2A1 (GLUT1) or GDF9. Results indicate a differential disruptive effect of mastitis induced by G- and G+ on oocyte developmental competence in association with alterations in maternal gene expression.

Naturally occurring mastitis disrupts developmental competence of bovine oocytes.

We examined the effects of naturally occurring mastitis on bovine oocyte developmental competence in vitro. Specifically, we investigated the effects of intramammary infection on the ovarian pool of oocytes (i.e., follicle-enclosed oocytes) and their ability to undergo in vitro maturation, fertilization, and further development to the blastocyst stage. Culled Holstein cows (n=50) from 9 commercial dairy farms in Israel were allotted to 3 groups according to somatic cell count (SCC) records of the last 3 monthly milk tests as well as of quarter samples collected before slaughter: (1) low SCC (n=7), (2) medium SCC (n=16), or (3) high SCC (n=27). Means of SCC values differed among low-, medium-, and high-SCC groups: 148,000, 311,000 and 1,813,000 cell/mL milk, respectively. Milk yield and days in milk did not differ among the 3 groups. Bacterial isolates included coagulase-negative staphylococci, Escherichia coli, Streptococcus dysgalactiae, or no bacteria found. Ovaries were collected at the abattoir and brought to the laboratory. Cumulus oocyte complexes were recovered separately from each cow and subjected individually to in vitro maturation and fertilization, followed by 8d in culture. The number of aspirated oocytes did not differ among groups, with a range of 17 to 21 oocytes per cow. The proportion of oocytes that cleaved into 2- to 4-cell-stage embryos (86.1 ± 3.4%) did not differ among groups. In contrast, mean percentages of embryos developed to the blastocyst stage on d 7 and 8 after fertilization were less in both medium- and-high SCC groups than in the low-SCC group (5.6 ± 2.3 and 4.1 ± 1.8 vs. 18.1 ± 4.6%, respectively). Additional analysis indicated that cleavage and blastocyst-formation rates did not differ among the bacterial types in the low-, medium-, and high-SCC groups. These are the first results to demonstrate that naturally occurring mastitis disrupts the developmental competence of the ovarian pool of oocytes, (i.e., oocytes at the germinal vesicle stage). The disruption was associated with elevation of SCC rather than bacterial type. The results may provide a partial explanation for the low fertility of cows that have contracted mastitic pathogens before insemination.

Real-time visual/near-infrared analysis of milk-clotting parameters for industrial applications.

The economical profitability of the dairy industry is based on the quality of the bulk milk collected in the farms, therefore it was based on the herd level rather than on the individual animals at real time. Udder infection and stage of lactation are directly related to the quality of milk produced on the herd level. However, improvement of milk quality requires testing each animal's milk separately and continuously. Recently, it was postulated that online equipment can estimate milk quality according to its clotting parameters, and thus result in better economical return for cheese making. This study further investigated the potential application of the AfiLab™ equipment to provide real-time analysis of milk-clotting parameters for cheese manufacture and cheese yield on quarter (1018) and individual cow (277) levels. Days in milk, lactose, log SCC and udder infection were found to have a significant effect on curd firmness and cheese properties and yield. The results clearly indicate that: (a) the parameter Afi-CF determined with the AfiLab™ is suitable for assessing milk quality for its clotting parameters, a value which is not provided by merely measuring fat and protein content on the gland and the cow levels; (b) bacterial type is the single major cause of reduced milk quality, with variations depending on the bacterial species; and (c) early and late lactation also had negative effects on milk-clotting parameters. Cheese made from the various milk samples that were determined by the Afilab™ to be of higher quality for cheese making resulted in higher yield and better texture, which were related mainly to the bacterial species and stage of lactation.

Lipopolysaccharide challenge of the mammary gland in cows induces nitrosative stress that impairs milk oxidative stability.

The aim of this work was to study the effects of mastitis induced by intramammary lipopolysaccharide (LPS) challenge on milk oxidative stability, as well as to understand the underlying biochemical processes that cause such changes. LPS challenge was associated with nitric oxide burst from the surrounding mammary epithelial cells and consequently induced nitrosative stress that was induced by the formation of NO2• from nitrite by lactoperoxidase. This response was associated with an ∼3-fold increased formation of hazardous compounds: nitrotyrosines, carbonyls and lipid peroxides. We sustained the involvement of xanthine oxidase as a major source of hydrogen peroxide. In consistent with previous findings, catalase has been shown to play a major role in modulating the nitrosative stress by oxidizing nitrite to nitrate. The current hygienic quality criteria cannot detect mixing of low-quality milk, such as milk with high somatic cells, and nitrite with high-quality milk. Thus, development of an improved quality control methodology may be important for the production of high-quality milk.

The effect of plasma removal from apheresis platelet concentrates on platelet function.

The effect of plasma removal on platelet function has scarcely been investigated. Plasma removal from apheresis platelet concentrates was achieved by centrifugation at 5000 g for 6 min or 2000 g for 10 min. After resting for 1 h, platelet concentrates were resuspended in 0·9% NaCl. Platelet function was tested before centrifugation and after resuspension by multiple electrode impedance aggregometry (MEA) and light transmission aggregometry (LTA). Plasma removal resulted in 10-14% lower response to TRAP-6 by MEA using both washing procedures, whereas TRAP-6-inducible aggregation by LTA increased slightly (2-5%). Neither plasma removal method affected collagen-induced aggregation. Thus, platelet function did not deteriorate significantly by either method.

Administration of prednisolone intravenously or dexamethason orally in random donors reveals equal results in granulocyte collection: a single centre experience.

Since cytokine--mobilization for granulocyte collections in random donors--is not licensed in Austria, we investigated in a prospective analysis the effects of prednisolone or dexamethasone (DXM) in 153 donors (35 women/118 men) and in the collected products. Either prednisolone (50 mg) intravenously 4 h before granulocyte collection or DXM (8 mg) orally 12 h before collection was given. Collections were performed with the Cobe Spectra (CardianBCT, Lakewood). We recorded a significantly higher percentage of granulocytes in the peripheral blood of the DXM. However, the collected number of granulocytes was equal in both groups.

Lipopolysaccharide challenge of the mammary gland in bovine induced a transient glandular shift to anaerobic metabolism.

Support of milk production in modern dairy cows demands a large proportion of its own metabolic resources, such as glucose, which might be required under stressful situations. The aim of the experiment was to test the hypothesis that acute immune stress shifts oxidative metabolism to glycolysis. Two mammary quarters in 6 Holstein cows were infused with lipopolysaccharide (LPS), whereas the 2 counter quarters served as controls to the treatment. An additional 6 cows were infused with saline and served as running controls. The LPS challenge induced dramatic transient increases in milk lactate (75-fold) and malate (11-fold) concentrations (both markers of glycolysis) at 24h posttreatment. No significant changes in lactate and malate concentrations were recorded in control quarters and control animals, indicating that the effect of LPS was restricted to the treated gland. The LPS challenge induced a dramatic transient decrease in milk yield, and lactose and citrate (a marker of mitochondrial metabolism) secretion at 24h posttreatment. The kinetics were inversely proportional to those of lactate and malate concentrations. Thus, our data suggest that LPS challenge induces acute conversion of epithelial cell metabolism from principally mitochondrial-oxidative to principally cytosolic (glycolytic), which allows the diversion of metabolic resources normally used to synthesize milk to support the immune system. An in vitro bacterial growth test showed that concentrations of lactate, malate, and lactose equivalent to those found in the in vivo experiment delayed and reduced the growth of a pathogenic Escherichia coli strain, suggesting that they play a role in diminution of bacterial multiplication in the mammary gland.

Association of conception rate with pattern and level of somatic cell count elevation relative to time of insemination in dairy cows.

The aim was to evaluate the effects of mastitis, determined by the pattern and level of somatic cell count (SCC) around first artificial insemination (AI), on conception rate (CR). Data from 287,192 first AI and milk records covering a 7-yr period were obtained from the Israeli Herd Book. Analyses examined the association of probability of conception with SCC elevation relative to timing of AI, using generalized linear mixed models. A SCC threshold of 150,000 cells/mL of milk was set to distinguish between uninfected cows and cows with mastitis. Accordingly, cows with high SCC before and low SCC after AI were designated cured, those with low SCC before and high SCC after AI were designated newly infected, and cows with high SCC before and after AI were designated chronic (likely subclinical) mastitic cows. Compared with uninfected cows, the cured, newly infected, and chronic subgroups showed reduced CR (39.4±0.1, 36.6±0.2, 32.9±0.3, and 31.5±0.2, respectively). In the chronic, subclinical group, probability of conception was lowered by 14.5% in the mild and moderately elevated SCC subgroups and by 20.5% in cows with high SCC elevation compared with the uninfected group (CR of 29.7 vs. 39.4%, respectively). A single high elevation of SCC (>10(6) cells/mL on only 1 milk test day) lowered the probability of conception by 23.6% when it occurred during the 10 d immediately before AI, but not when it occurred earlier. For 30 d after AI, probability of conception was lowered by about 23%, as reflected in a CR of about 27% compared with the uninfected group. Probability of conception was lowered in cows with uterine and foot health problems (33.9%), in multiparous cows (34.1%), and in cows in the summer (29.1%), but no interactions with mastitis were detected. Results indicate that SCC elevation around AI, typical for subclinical mastitis, was associated with a significant reduction in probability of conception, and that even mild SCC elevation reduced CR. Severe elevation of SCC before AI, typical for clinical intramammary infection, reduced the probability of conception.