Effect of stage of lactation and parity on mammary gland cell renewal

Milk production is a function of the number and activity of mammary epithelial cells, regardless of stage of lactation. Milk yield is generally higher in multiparous cows than in primiparous cows, but persistency is usually greater in the latter group. We compared several measures related to metabolic activity, apoptosis, and endocrine control of mammary cell growth in 8 primiparous and 9 multiparous cows throughout lactation. Mammary gland biopsies were taken in early [10 d in milk (DIM)], peak (50 DIM), and late (250 DIM) lactation to evaluate gene expression and determine DNA and fatty acid synthase (FAS) content. Milk samples taken the day before the biopsies were used to detect protease activities and to determine stanniocalcin-1 (STC) concentrations. Blood samples served to measure insulin-like growth factor-1, prolactin, and STC concentrations. Milk yield was higher in multiparous cows than in primiparous cows at the 10 DIM (32.8 ± 1.3 and 25.2 ± 0.8 kg/d) and 50 DIM (38.0 ± 1.2 and 29.8 ± 1.1 kg/d), but it was the same for both groups at 250 DIM (23.9 ± 1.5 and 23.8 ± 1.1 kg/d). Except for stearoyl-coenzyme A desaturase, expression of genes related to milk synthesis was not affected by stage of lactation. However, gene expression of acetyl-coenzyme A carboxylase, β-casein, and FAS was lower in early lactation in primiparous cows. Expression of both proapoptotic bax and antiapoptotic bcl-2 genes was higher in primiparous cows, whereas the bax-to-bcl-2 ratio was not changed. Mammary DNA concentration was higher in multiparous cows, as was the amount of FAS protein in early lactation. Two bands of protease activity were found in milk samples, and one of the bands had an apparent molecular weight similar to gelatinase A and was dependent on the stage of lactation. Serum insulin-like growth factor-1 increased with day of lactation and was higher in primiparous cows. Serum prolactin decreased in late lactation, but peak values were observed in early lactation for primiparous cows and peak lactation for multiparous cows. Milk STC content increased with advancing lactation. The results are consistent with a lower degree of differentiation and a greater capacity for cell renewal in the mammary gland of primiparous cows.     

Factors affecting the lactoferrin concentration in bovine milk

Lactoferrin (LF) concentrations in the milk with different levels of the somatic cell count score were examined using an ELISA to determine whether milk LF concentration is influenced by parity of the cow, stage of lactation, and the somatic cell count. The study animals were 198 Chinese Holstein cows randomly chosen from more than 1,600 cows in 4 dairy farms in the Beijing area. The cows had shown no sign of mastitis for 2 mo. Daily milk production was recorded, and milk samples were taken from individual cow samples. The LF concentration varied between 31.78 and 485.63 μg/mL in milk from normal animals. Lactoferrin was significantly associated with stage of lactation (r = 0.557) and daily milk production (r = −0.472). Nevertheless, there was no significant relationship with parity. Moreover, milk LF concentration tended to be correlated with the somatic cell count score (r = 0.375). This finding suggests that milk LF may be helpful as an indicator for intramammary infection in dairy cows.     

Fresh cow mastitis monitoring on day 3 postpartum and its relationship to subsequent milk production

The purpose was to determine the association of milk California Mastitis Test (CMT), somatic cell concentration (SCC), and milk differential cell count results on day 3 postcalving with subsequent lactation production and health events. On d 3 postcalving, the CMT was performed and quarter milk samples were collected from 130 dairy cows. Quarter SCC and milk differential cell counts were determined. Microbiology on duplicate quarter milk samples was used to determine the presence of intramammary infection by major or minor pathogens. Production measures obtained using Dairy Herd Improvement Association testing were 150-d standardized and summit milks. Milk culture results on a cow basis included 82 (63.1%) samples with no growth, 31 (23.9%) with major pathogens, and 17 (13.1%) with minor pathogens. Milk culture results comparing cows with no growth to those with any growth (major or minor pathogens) were not associated with statistically significant differences in milk production. Milk culture results comparing cows with major pathogens to those with no growth and minor pathogens combined were associated with statistically significant differences in 150 d milk. Milk production did not differ for cows with CMT results above and below a cut-off of trace, and for SCC results above and below cut-offs of 200,000, 300,000, and 400,000/mL, respectively. Statistically significant differences in milk production were found for cows above and below cut-offs for percentage neutrophils in milk and for absolute neutrophil counts. Associations were found for milk production and number of quarters (0, 1, 2, or 3 and 4 combined) above respective cut-offs for SCC, percentage neutrophils in milk, and absolute numbers of neutrophils in milk, but not for CMT. Milk production differed for cows experiencing any health event versus those with no health event. The most commonly recorded health event was clinical mastitis. Statistically significant associations were detected between health events and milk culture results, SCC, neutrophil percentage, and neutrophil absolute counts. Results of the present investigation indicate that milk monitoring on d 3 of lactation using milk neutrophil percentage or neutrophil absolute counts may be useful as an indication of subsequent milk production.     

Effects of Parity,Bacteriological Status,Stage of Lactation,and Dry Period on N-Acetyl-β-D-Glucosaminidase Activity of Milk and Dry Secretion

Quarter foremilk samples from 61 cows were obtained at 1.5, 3, 21, and 35 wk of lactation and at 7 d after drying off. Measurements for each sample were milk SCC, NAGase activity in the milk and ability of milk to promote phagocytosis of Staphylococcus aureus by polymorphonuclear leukocytes. Milk SCC and NAGase were correlated (r = .62). The NAGase in dry cow secretion was 10-fold higher than in milk. Parity differences in NAGase activity were not significant. There were large stage of lactation trends in NAGase: NAGase activity was high in early lactation, decreased in midlactation, and increased in late lactation and in dry secretion. The increase in activity of the enzyme in milk of first parity cows in late lactation was not as great as in milk of second and third lactation cows. N-Acetyl-β-D-glucosaminidase activity of milk from quarters with intramammary infections was higher than that of milk from quarters free of pathogens.     

Experimental challenge of bovine mammary glands with Enterococcus faecium during early and late lactation

Mammary glands of early and late lactation cows were challenged with Enterococcus faecium of bovine origin to determine in vivo pathogenicity and milk somatic cell count (SCC) responses. A total of 20 early lactation and 18 late lactation mammary glands were challenged. Two isolates highly adaptive and 2 isolates poorly adaptive for in vitro growth in mammary secretion were used as challenge strains of bacteria. Challenged quarters of early lactation cows were more susceptible to intramammary infection caused by E. faecium than those of late lactation cows. Intramammary challenge with isolates poorly adaptive for in vitro growth in mammary secretions resulted in 94.7% of quarters infected compared with 36.8% of the quarters infused with the isolates highly adaptive for in vitro growth in mammary secretions. Milk from quarters infused with the isolates poorly adaptive for in vitro growth had higher SCC and bacterial counts compared with quarters challenged with the isolates highly adaptive for in vitro growth. A stage of lactation effect within treatment groups was measured when milk SCC were compared between early and late lactation cows. Milk SCC in uninfused (negative control) quarters were lower in early lactation cows compared with late lactation cows. Conversely, in quarters infused with isolates poorly adaptive for in vitro growth, SCC were higher in early lactation cows compared with late lactation cows on d 2, 3, 4, 15, 16, and 17 postchallenge. In quarters infused with isolates highly adaptive for in vitro growth, SCC response did not differ between early and late lactation cows. In vitro growth of E. faecium in mammary secretion was inversely related to in vivo pathogenicity in the mammary glands of early and late lactation cows.     

Effect of continuous milking and bovine somatotropin supplementation on mammary epithelial cell turnover

Objectives were to determine effects of continuous milking (CM) and bovine somatotropin (bST) administration on 1) mammary epithelial cell (MEC) proliferation, apoptosis, and ultrastructure during late gestation and early lactation, 2) expression of genes associated with proliferation, and apoptosis in mammary epithelial cells, and 3) milk yield and composition. Second-gestation, first dry-period cows were randomly assigned to either continuous bST throughout late gestation and early lactation (+bST; n = 4) or no bST (−bST; n = 4) administration. Within each animal, udder halves were randomly assigned to CM or a 60-d dry period (control) treatment. Daily milk yield and weekly milk composition were measured during the last 60 d of gestation in CM halves and from 1 to 30 d postpartum for both halves. Mammary biopsies were obtained at −20 ± 7, −8 ± 3, +1 ± 0, +7 ± 0, and +20 ± 0 d (mean ± standard error) relative to parturition. Prepartum half-udder milk yield was greater in +bST cows than in −bST cows (9.9 vs. 8.2 kg/d) and postpartum half-udder milk yields were dramatically reduced in CM halves compared with control halves (10.6 vs. 22.2 kg/d), regardless of bST treatment. Proliferation of MEC was reduced in CM halves at −8 d (2.7 vs. 5.4%). Apoptosis of MEC was elevated during early lactation for d +1 and +7 in control halves, but was only increased at d +1 in CM halves. Turnover of MEC was not affected by bST. Ultrastructure data indicated complete involution of the control half and lactation maintenance in CM glands (d −20). By d −8, control tissue contained alveoli in an immature secretory state, but CM tissue contained both lactating and immature alveoli. Postpartum ultrastructure parameters were similar between halves until d 20 when control tissue was composed of a homogeneous population of lactating alveoli, but CM tissue contained lactating, engorged, and resting alveoli. Expression of CCAAT/enhancer binding protein-β (CEBP-β), cyclin D1, and bcl₂ were up-regulated during late gestation, but did not differ between control and CM halves. Expression of α-lactalbumin was increased in CM halves during late gestation, but was not different in CM and control tissue after parturition. Other genes evaluated (bax, insulin-like growth factor binding protein 5, ATP-binding cassette 1, and p27) were not differentially expressed at any timepoints evaluated. Results indicate that CM reduced subsequent half-udder milk yield in primiparous cows through altered MEC turnover and secretory capacity. Negative effects of CM on the subsequent lactation were not alleviated by bST supplementation.     

Lactation failure in crossbred Sahiwal Friesian cattle

Milk producers in Malaysia make extensive use of crossbred Sahiwal Friesian dairy cattle. These animals have, however, been found susceptible to lactation failure. A survey of cows in an experimental herd of F1 Sahiwal Friesian animals indicated that, in 30% of animals, milk yield decreased to negligible levels within the first 8 weeks post partum. Lactation failure was associated with a progressive increase in the amount of residual milk left in the udder after normal milking. By week 3 of lactation, residual milk volume was significantly greater than that in animals that, based on previous lactation history were not susceptible to lactation failure, and accounted for up to 30% of milk available at the morning milking. The cellular consequences of residual milk accumulation were evident in the activities of acetyl-CoA carboxylase, fatty acid synthetase and galactosyltransferase, key enzyme markers of cellular differentiation, which decreased in glands undergoing lactation failure and were lower than values measured in tissue of control cows. Mammary cell number, estimated by tissue DNA content, was also reduced in animals undergoing lactation failure. These indices of mammary development indicate that lactation failure is the result of premature involution in susceptible animals. Premature involution is a predictable consequence of progressive milk stasis in failing lactation, and attributable to an increase in autocrine feedback by inhibitory milk constituents. The progressive increase in residual milk is, on the other hand, unlikely to be attributable to impaired mammary development. Measurements of milk storage during milk accumulation showed no differences between control and lactation failure cows in the distribution of milk between alveolar and cisternal storage compartments. We conclude that lactation failure in Sahiwal Friesian cows is due to a failure of milk removal, and probably the result of an impaired milk ejection reflex rather than to the glands' milk storage characteristics.     

Effects of 6 times daily milking during early versus full lactation of Holstein cows on milk production and blood metabolites

An experiment was conducted to determine the effects of different milking frequencies on entire lactation production performance in Holstein cows. One hundred twenty Holstein cows were assigned to 3 milking treatments (35 multiparous and 5 primiparous cows in each): 1) milking 6 times daily for the entire lactation (6×); 2) milking 6 times daily for the first 90 d in milk (DIM) and switching to 3 times daily milking afterward (6×-3×); and 3) milking 3 times daily for the entire lactation (3×). Milk yield was recorded every other day during the first 60 DIM and on 2 consecutive days per week subsequently. Cows were weighed and scored for their body condition immediately after parturition and monthly afterward. Blood and milk samples were taken from each cow on 30, 60, 90, 120, 150, 210, and 270 DIM, with an additional blood sampling on 15 DIM. Milk and fat corrected milk yield was greater for 6× and 6×-3× cows than for 3× cows (36.82, 37.32, and 36.1, 36.75 versus 34.56, 35.33 kg/d, respectively) during the experimental period. Milk fat and lactose percentage were not different among treatments, but milk protein was lower in 6× cows than in 6×-3× and 3× cows. Blood glucose concentration was higher and blood nonesterified fatty acids and β-hydroxybutyrate concentration were lower in 3× cows than in 6× and 6×-3× cows during early lactation. Dry matter intake was greater in 6× cows than in 6×-3× and 3× cows (23.05, 22.58, and 22.45 kg/d, respectively). The 3× cows began to gain weight earlier than the 6× and 6×-3× cows, but there was no difference among groups for BW change regarding the entire experimental period. These results indicate that increasing milking frequency to 6 times daily increases milk yield only during early lactation and that there are no advantages of milking 6 times daily compared with 3 times daily during the mid and late lactation periods. Given the results of this study and the economical aspects of production, milking 6 times daily until 90 DIM and subsequently switching to milking 3 times daily is preferred.     

Effect of a mixture of supplemental dietary plant essential oils on performance of periparturient and early lactation dairy cows

Plant essential plant oils (EO) are volatile aromatic compounds with antimicrobial activity that can alter ruminal fermentation when used as dietary supplements. A feeding trial was conducted to determine the effects of dietary supplementation of periparturient and early lactation dairy cows with a specific mixture of EO. Forty multiparous Holstein cows were randomly assigned to either control (C) or EO-supplemented (1.2 g/cow per day) total mixed rations (TMR). Feeding of treatment diets commenced 3 wk before the expected calving date and continued through 15 wk in lactation. The prepartum TMR contained 70% forage [70% corn silage, 15% alfalfa silage, and 15% wheat straw; dry matter (DM) basis]. The lactation TMR contained 50% forage (60% corn silage, 33% alfalfa silage, 7% alfalfa hay; DM basis). Prepartum and lactation TMR were formulated to contain 12 and 17% CP (DM basis), respectively. There were no differences between treatments for prepartum DM intake (DMI), but DMI was 1.8 kg/d less for EO than C on average across the 15-wk lactation trial. Plasma concentrations of glucose, nonesterified fatty acids, β-hydroxybutyrate, and urea-N on samples collected −21, −14, −7, −1, 1, 8, 15, 22, and 29 d relative to calving were unaffected by treatment. There were no differences between treatments for actual or fat-corrected milk yields on average across the 15-wk lactation trial. Milk protein content was 0.15% units less for EO than C. Feed efficiency (kg of milk per kg of DMI) tended to be greater for EO than C on average and was greater during wk 8 to 14 of lactation. Prepartum and lactation body weight and condition score measurements were unaffected by treatment. There was no benefit to EO in prepartum dairy cows. Dietary supplementation with EO reduced DMI in early lactation dairy cows with no effect on milk yield.     

Evaluation of milk enzymes and electrolytes, plasma metabolites, and oxidative status in twin cows milked in an automatic milking system or twice daily in a conventional milking parlor

The aim of this paper was to evaluate the effects of automatic milking (AM) on milk enzymes and minerals related to mammary epithelial integrity in comparison with twice-daily conventional milking (CM). One cow from each of 6 pairs of twins was assigned to be milked with AM or with CM throughout first lactation. Milk production was recorded and milk samples were collected at 4, 11, 18, 25, 32, and 39 wk of lactation (WOL) to determine fat and protein content, somatic cell count, pH, plasminogen (pl) and plasmin (Pl) activities, Na, K, and Cl. Body condition score was monitored; blood samples were collected to determine energy-related metabolites in the first third of lactation (14 WOL), and plasma oxidative status throughout lactation. Overall mean and standard deviation of milking frequency (MF) in AM were 2.69 and 0.88, respectively. Milk production, fat and protein contents, and somatic cell count did not differ between milking systems. The pl and pl+Pl activities were lesser in AM than in CM. Milk pH was greater in AM than in CM. Milk Na, K, Na/K ratio, and Cl did not differ across the whole lactation. Milk pH had a positive correlation with milk Pl activity (r = 0.41), Na (r = 0.37), and Cl (r = 0.40) concentration, and negative correlation with the log₁₀ of pl/Pl ratio (r = −0.47). The milk Na/K ratio had a positive correlation (r = 0.55) with milk Pl activity. Milking system (MS) did not seem to affect mammary epithelial permeability. The differences in enzymatic (proteolytic) activity due to the MS, probably related to daily MF, lead one to suppose that the quality of the protein fraction for the cheese-making process was preserved better with AM than with CM, even if differences in pH might negatively interfere. No difference was detected in BCS, and in plasma concentration of triglycerides and nonesterified fatty acids, whereas plasma cholesterol concentration during the first 10 WOL was lesser in AM than CM. Oxidative status, measured by plasma reactive oxygen metabolites and thiol groups, did not differ between MS throughout the whole lactation. These results suggest that early lactation of AM primiparous cows may give rise to crucial situations: for milk production, when a low MF may impair further mammary cell proliferation; for milk quality, if an irregular MF, with prolonged milking intervals, leads to an increased milk pH with increased conversion of pl to Pl.     

Effect of heat stress during the dry period on mammary gland development

Heat stress during the dry period negatively affects hepatic metabolism and cellular immune function during the transition period, and milk production in the subsequent lactation. However, the cellular mechanisms involved in the depressed mammary gland function remain unknown. The objective of the present study was to determine the effect of heat stress during the dry period on various indices of mammary gland development of multiparous cows. Cows were dried off approximately 46 d before expected calving and randomly assigned to 2 treatments, heat stress (HT, n = 15) or cooling (CL, n = 14), based on mature equivalent milk production. Cows in the CL treatment were provided with sprinklers and fans that came on when ambient temperatures reached 21.1°C, whereas HT cows were housed in the same barn without fans and sprinklers. After parturition, all cows were housed in a freestall barn with cooling. Rectal temperatures were measured twice daily (0730 and 1430 h) and respiration rates recorded at 1500 h on a Monday-Wednesday-Friday schedule from dry off to calving. Milk yield and composition were recorded daily up to 280 d in milk. Daily dry matter intake was measured from dry off to 42 d relative to calving. Mammary biopsies were collected at dry off, −20, 2, and 20 d relative to calving from a subset of cows (HT, n = 7; CL, n = 7). Labeling with Ki67 antigen and terminal deoxynucleotidyl transferase deoxyuridine triphosphate nick-end labeling were used to evaluate mammary cell proliferation and apoptosis, respectively. The average temperature-humidity index during the dry period was 76.6 and not different between treatments. Heat-stressed cows had higher rectal temperatures in the morning (38.8 vs. 38.6°C) and afternoon (39.4 vs. 39.0°C), greater respiration rates (78.4 vs. 45.6 breath/min), and decreased dry matter intake (8.9 vs. 10.6 kg/d) when dry compared with CL cows. Relative to HT cows, CL cows had greater milk production (28.9 vs. 33.9 kg/d), lower milk protein concentration (3.01 vs. 2.87%), and tended to have lower somatic cell score (3.35 vs. 2.94) through 280 d in milk. Heat stress during the dry period decreased mammary cell proliferation rate (1.0 vs. 3.3%) at −20 d relative to calving compared with CL cows. Mammary cell apoptosis was not affected by prepartum heat stress. We conclude that heat stress during the dry period compromises mammary gland development before parturition, which decreases milk yield in the next lactation.     

Factors affecting ability of skim milk to support phagocytosis by bovine polymorphonuclear leukocytes

Milk samples from quarters were collected from 48 Holstein cows at 1.5, 3, 21, and 35 wk postpartum and at 1 wk after drying off. Foremilk samples were obtained for bacteriological examination, somatic cell count of milk, and for assay of phagocytosis. Thirty-two cows in first lactation and 16 in third lactation produced by two criteria of sire selection (high milk versus multiple trait) were studied. In vitro phagocytosis was assayed four times on each original quarter milk sample, yielding 3,840 determinations. Variance components were large for cow-associated variation. Cow, stage X cow, and cow X sample dilution were 18.7, 20.6, and 6.0% of total variance. Phagocytosis was 9% higher in skim milk from third lactations than from first. Phagocytosis was highest in dry period samples, followed by 1.5, 35, 3, and 21-wk postpartum samples. Milk somatic cell count tended to be related more closely to phagocytosis than was current bacteriological status of the quarter. Skim milk from genetically higher producing cows was less conducive to phagocytosis than skim milk from genetically lower producing cows, possibly because of dilution.     

Effect of somidobove sustained release administration on the lactation performance of dairy cows

Lactation performance was determined on 190 multiparous Holsteins from five herds supplemented with 0, 320, 640, or 960 mg of somidobove every 28 d. The experiment consisted of 21 d of pretreatment and treatment periods of various lengths, depending upon stage of lactation of animals at first administration. Somidobove beginning in early (28 to 45 d in milk), mid (111 to 166 d in milk), or late (166 to 334 d in milk) stages of lactation consisted of 9, 6, or 3 administrations. Milk and 3.5% FCM yields were increased by each dose of somidobove in all stages. Milk composition and dry matter and energy intakes were similar among treatments within stage. Milk to DMI ratio and milk energy to net energy intake ratio were improved by somidobove. Gain was positive for all treatments, but less in somidobove-supplemented cows. Lower body weight and condition score at the completion of somidobove treatment resulted. For early cows, days to first estrus and days to first breeding were similar; however, total number of inseminations for cows receiving somidobove was twofold greater than control, resulting in a longer calving interval. Results demonstrated efficacy of somidobove administered every 28 d to lactating dairy cattle for increased milk yield.     

Effects of stage of lactation and time of year on plasmin-derived proteolytic activity in bovine milk in New Zealand

The objective of this study was to determine the effects of stage of lactation (SOL) and time of year on plasmin-derived proteolytic activity in the milk of pasture-fed dairy cows in New Zealand. Four herds of 20 Friesian cows were used, one herd calving in each of January, April, July and October. Cows grazed ryegrass/white clover pasture only, except during June (winter) when all cows received supplementary pasture silage. Milk samples were collected on four occasions during the year (spring, summer, autumn and winter) from each cow in milk, to give a total of three samples per cow (early, mid and late lactation; c. 30, 120 and 220 days after calving, respectively). Milk samples were analysed for plasmin-derived proteolytic activity. There was no effect of either SOL or time of year on plasmin activity and therefore yields of plasmin followed patterns in milk yield (highest in early lactation and in summer). There were effects of both SOL and time of year on plasminogen-derived and total plasmin plus plasminogen-derived activity, both of which were highest in late lactation and in spring. Changes in plasminogen-derived activity and total plasmin plus plasminogen-derived activity due to SOL were not only due to the decrease in milk yield associated with advancing lactation, because enzyme yields were also increased with advancing lactation. Similarly, effects of time of year on plasminogen-derived activity and total plasmin plus plasminogen-derived activity could not be attributed solely to concomitant changes in milk yield, and may be influenced by the variation in the quality and quantity of feed during the year inherent in a pasture-based dairy system. Effects of SOL on proteolytic activity were greater than, and independent of, effects of time of year.     

Genetic merit for fertility traits in Holstein cows: IV. Transition period,uterine health,and resumption of cyclicity

The objective of this study was to monitor the dry matter intake (DMI), metabolic status, uterine health, and resumption of cyclicity in cows with similar genetic merit for milk production traits but with either good (Fert+) or poor genetic merit (Fert−) for fertility traits. Twenty-six cows were enrolled in the study and data are reported for 15 Fert+ and 10 Fert− cows that completed the study. All cows received a total mixed ration diet during early lactation and were turned out to pasture in late spring. Dry matter intake was recorded daily from wk −2 to 5 relative to parturition. Blood metabolites and metabolic hormones were measured from wk −2 to 8 relative to parturition. Milk production, body condition score, and body weight until wk 35 of lactation are reported. To monitor uterine health, vaginal mucus was scored weekly on a scale of 0 (no pus) to 3 (≥50% pus) from parturition to wk 8 and uterine polymorphonuclear neutrophil count was measured at wk 3 and 6 postpartum. Prepartum DMI was similar between genotypes, but Fert+ cows had significantly greater DMI than Fert− cows (19.7 vs. 16.8 kg of dry matter/d) during the postpartum period. Energy balance at wk 1 was significantly greater in Fert+ cows than in Fert− cows [2.3 vs. −1.12 unité fourragère lait (UFL)/d]. The Fert+ cows had significantly greater daily milk solids production (1.89 vs. 1.74 kg/d) and tended to have greater daily milk yield (24.2 vs. 22.3 kg/d). The Fert+ cows had significantly greater mean circulating insulin-like growth factor-I (102.62 vs. 56.85 ng/mL) and tended to have greater mean circulating insulin (3.25 vs. 2.62 μIU/mL) compared with Fert− cows from wk −2 to 8 relative to parturition. Mean circulating glucose (3.40 vs. 3.01 mmol/L) concentrations were significantly greater in Fert+ cows compared with Fert− cows from wk −2 to 3 relative to parturition. The Fert+ cows maintained significantly greater mean body condition score throughout lactation compared with Fert− cows (2.98 vs. 2.74 units). Moreover, Fert+ cows had better uterine health compared with Fert− cows, as evidenced by lower weekly vaginal mucus scores from wk 2 to 6 postpartum and, based on uterine cytology, smaller proportions were classified as having endometritis at wk 3 (0.42 vs. 0.78) and 6 (0.25 vs. 0.75). Also, a significantly greater proportion of Fert+ cows had resumed cyclicity by wk 6 postpartum (0.86 vs. 0.20) compared with Fert− cows. Hence, we report for the first time that genetic merit for fertility traits is associated with postpartum uterine health status. Superior uterine health and earlier resumption of cyclicity may be mediated through differences in DMI, energy balance, insulin, insulin-like growth factor-I, and body condition score profiles. Importantly, phenotypic improvement in fertility traits was achieved without antagonizing milk production.     

Dietary cation-anion difference and dietary protein effects on performance and acid-base status of dairy cows in early lactation

Our objective was to examine the effects of dietary cation-anion difference (DCAD) with different concentrations of dietary crude protein (CP) on performance and acid-base status in early lactation cows. Six lactating Holstein cows averaging 44 d in milk were used in a 6 × 6 Latin square design with a 2 × 3 factorial arrangement of treatments: DCAD of −3, 22, or 47 milliequivalents (Na + K - Cl - S)/100 g of dry matter (DM), and 16 or 19% CP on a DM basis. Linear increases with DCAD occurred in DM intake, milk fat percentage, 4% fat-corrected milk production, milk true protein, milk lactose, and milk solids-not-fat. Milk production itself was unaffected by DCAD. Jugular venous blood pH, base excess and HCO₃⁻ concentration, and urine pH increased, but jugular venous blood Cl⁻ concentration, urine titratable acidity, and net acid excretion decreased linearly with increasing DCAD. An elevated ratio of coccygeal venous plasma essential AA to nonessential AA with increasing DCAD indicated that N metabolism in the rumen was affected, probably resulting in more microbial protein flowing to the small intestine. Cows fed 16% CP had lower urea N in milk than cows fed 19% CP; the same was true for urea N in coccygeal venous plasma and urine. Dry matter intake, milk production, milk composition, and acid-base status did not differ between the 16 and 19% CP treatments. It was concluded that DCAD affected DM intake and performance of dairy cows in early lactation. Feeding 16% dietary CP to cows in early lactation, compared with 19% CP, maintained lactation performance while reducing urea N excretion in milk and urine.     

Clinical disease and stage of lactation influence shedding of Mycobacterium avium subspecies paratuberculosis into milk and colostrum of naturally infected dairy cows

Mycobacterium avium ssp. paratuberculosis (MAP) is the causative agent of Johne's disease (JD). One mode of transmission of MAP is through ingestion of contaminated milk and colostrum by susceptible calves. The objective of this study was to determine if the amount of MAP shed into the milk and colostrum of infected cows was affected by severity of infection as well as the number of days in milk (DIM). Milk was collected over the 305-d lactation period from naturally infected cows in the asymptomatic subclinical (n = 39) and symptomatic clinical (n = 29) stages of disease, as well as 8 noninfected control cows. All milk samples were assayed for MAP by culture on Herrold's egg yolk medium and either BACTEC 12B (Becton Dickinson, Franklin Lakes, NJ) or para-JEM (Thermo Fisher Scientific, Trek Diagnostic Systems Inc., Cleveland, OH) liquid medium, and by direct PCR for the IS900 target gene. Mycobacterium avium ssp. paratuberculosis was detected in 3.8, 4.1, and 12.6% of milk samples collected from cows with subclinical JD after culture in Herrold's egg yolk medium, liquid medium, and direct PCR, respectively. The frequency of MAP positivity increased to 12.9, 18.4, and 49.2% of milk samples collected from cows with clinical JD by these same methods, respectively. None of the milk samples collected from control cows was positive for MAP by any detection method. Viable MAP was primarily isolated from milk and colostrum of subclinically and clinically infected cows collected in early lactation (DIM 0–60), with negligible positive samples observed in mid (DIM 60–240) and late (DIM 240–305) lactation. This study demonstrates that shedding of MAP into milk is affected by infection status of the cow as well as stage of lactation, providing useful information to producers to help break the cycle of infection within a herd.     

Local control of mammary involution: Is stanniocalcin-1 involved?

There is considerable evidence to indicate the existence of local control of mammary gland involution, but the exact nature of this control has yet to be defined. Stanniocalcin-1 (STC-1) is a newly discovered mammalian hormone that seems involved in the lactation process and may be implicated in the control of involution. As a first step in investigating this hypothesis, the change in STC-1 levels in milk and serum was measured during drying off. Nine Holstein cows in late lactation were milked twice daily on half the gland, while the other half was left unmilked for a 14-d period. Milk and blood samples and mammary biopsies were taken on d −7, 1, 2, 7, and 14 relative to the onset of the nonmilking period. The concentrations of STC-1 in serum and milk were determined by RIA. The albumin concentration and proteinase activity of the milk were determined. Apoptosis of the mammary epithelium was quantified by terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling assay. Finally, the effects of milk on cellular activity and apoptosis were tested in vitro on mammary epithelial cells by measuring the turnover of tetrazolium salts and by counting the terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling-positive cells. The drying off of 2 quarters increased the milk production of the quarters that were milked by 30%. Milk proteinase activity and BSA and STC-1 concentrations increased in the nonmilked quarters, but remained unchanged in the milked quarters. Moreover, at d 2, the apoptotic rate of the mammary cells was higher in the nonmilked quarters than in the milked quarters (0.22 ± 0.04 vs. 0.07 ± 0.04%, respectively). Finally, in vitro experimentation demonstrated that mammary epithelial cells cultured in the presence of milk from involuting quarters had 3-fold more apoptotic cells as compared with those cultured in milk from the milked quarters at d 14. The metabolic rate was reduced by 14.6% for milk from d 7 and 23.6% for milk from d 14. Interestingly, the metabolic rate was negatively correlated with the STC-1 concentration in milk (r = −0.65). This study shows for the first time that STC-1 in milk is increased following milk stasis, although its exact role in the involution process remains to be clarified.     

Prediction of ammonia emission from dairy cattle manure based on milk urea nitrogen: relation of milk urea nitrogen to urine urea nitrogen excretion

The objectives of this study were to assess the relationship between urinary urea N (UUN) excretion (g/d) and milk urea N (MUN; mg/dL) and to test whether the relationship was affected by stage of lactation and the dietary crude protein (CP) content. Twelve lactating multiparous Holstein cows were randomly selected and blocked into 3 groups of 4 cows intended to represent early [123 ± 26 d in milk (DIM); mean ± standard deviation], mid (175 ± 3 DIM), and late (221 ± 12 DIM) lactation stages. Cows within each stage of lactation were randomly assigned to a treatment sequence within a split-plot Latin square balanced for carryover effects. Stage of lactation formed the main plots (squares) and dietary CP levels (15, 17, 19, and 21% of diet dry matter) formed the subplots. Graded amounts of urea were added to the basal total mixed ration to linearly increase dietary CP content while maintaining similar concentrations of all other nutrients among treatments. The experimental periods lasted 7 d, with d 1 to 6 used for adjustment to diets and d 7 used for total collection of urine as well as milk and blood sample collection. Dry matter intake and yields of milk, fat, protein, and lactose declined progressively with lactation stage and were unaffected by dietary CP content. Milk and plasma urea-N as well as UUN concentration and excretion increased in response to dietary CP content. Milk and urine urea-N concentration rose at increasing and decreasing rates, respectively, as a function of plasma urea-N. The renal urea-N clearance rate differed among lactation stages and dietary CP contents. The relationship between UUN excretion and MUN differed among lactation stages and diverged from linearity for cows in early and late lactation. However, these differences were restricted to very high MUN concentrations. Milk urea N may be a useful tool to predict the UUN excretion and ultimately NH₃ emission from dairy cattle manure.     

Effect of dry period management on mammary gland function and its endocrine regulation in dairy cows

The objective of this study was to evaluate the effect of shortening the dry period on the mammary gland and the hormonal regulation of its functions. Holstein cows (n = 18) were assigned to a short dry period (SDP; 35 d; n = 9) or a conventional dry period (CDP; 65 d; n = 9). All cows were fed the same diets, with the exception that, during the dry period, the SDP cows received only the pre-calving diet for 35 d, whereas the CDP cows were fed a high-fiber diet from 65 to 28 d before calving and then received the same pre-calving diet as the SDP cows. Mammary gland functional capacity was evaluated at 70 days in milk, and mammary biopsies were taken in early and midlactation. Dry period length averaged 64.3 ± 1.1 and 31.9 ± 1.0 d for the CDP and SDP cows, respectively. The SDP cows had a lower milk yield and a lower energy-corrected milk yield compared with the CDP cows. The SDP cows also had a lower dry matter intake from wk 5 to 20 of lactation and tended to have lower plasma concentrations of β-hydroxybutyrate from wk 1 to 4. Prepartum serum progesterone and estradiol concentrations were unaffected by the dry period management. Serum growth hormone concentrations and milking-induced prolactin release were similar in both groups. However, during the period when the CDP cows were dry but the SDP cows were still being milked (wk −9 to −6), serum prolactin concentrations were higher in the SDP cows than in the CDP cows. The SDP cows had a lower milk BSA content than the CDP cows after the dry period and similar milk lactose concentrations, suggesting that their mammary tight junctions were closed following parturition and, therefore, that the later stage of their lactogenesis was not impaired by SDP management. In early and midlactation, mammary cell apoptosis and proliferation rates as well as mammary expression of genes involved in the function of this tissue were unaffected by the dry period management strategy. For cows in their second lactation, mammary gland functional capacity at 70 d in milk tended to be lower in the SDP cows. In conclusion, even though SDP management decreased milk production during the subsequent lactation, it did not affect mammary cell activity. Although direct evidence is still lacking, decreased mammary cell growth during the dry period is likely responsible for this negative effect. The higher prolactin concentrations in lactating cows during late gestation could be involved in this effect. More research is needed to test these hypotheses.