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-beta (CEBP-beta), cyclin D1, and bcl(2) were up-regulated during late gestation, but did not differ between control and CM halves. Expression of alpha-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.

The objective of this study was to determine the effects of altering dry period length in multiparous dairy cows (n = 341) on milk production for a full lactation (294 d). The study used 3 commercial herds in the western United States. Cows producing greater than 8,400 kg of mature-equivalent milk were assigned to treatments 60 d before their due dates. The 4 treatments were 1) 60-d dry period, label use of recombinant bovine somatotropin (rbST; 60d-L); 2) 32-d dry period, label use of rbST (32d-L); 3) 0-d dry period, label use of rbST (0d-L); and 4) 0-d dry period, continuous use of rbST (0d-C). Cows with shortened dry periods produced 625, 1,000, and 1,042 kg of milk during the prepartum period for treatments 2 to 4, respectively. During the postpartum period, cows on the 32d-L treatment produced similar amounts of milk compared with the 60d-L treatment (11,490 vs. 11,968 kg, respectively). However, cows on the 0d-L (10,316 kg) and 0d-C (10,195 kg) treatments produced significantly lower amounts of milk during the postpartum period compared with the 60d-L treatment. Total milk production from the prepartum and postpartum periods was not altered significantly and was 11,974, 12,112, 11,310, and 11,230 kg for treatments 1 to 4, respectively. The concentrations of beta-hydroxybutyrate and nonesterified fatty acids in serum after calving were decreased for cows on the 32d-L, 0d-L, and 0d-C treatments compared with cows on the 60d-L treatment, which may indicate improved metabolic status.

Aims:  To examine the variability in faecal shedding of Salmonella and Escherichia coli O157:H7 in healthy lactating dairy cattle and to evaluate the genetic relatedness of Salmonella isolates. Methods:  Faecal samples were obtained from lactating Holstein dairy cattle on four commercial farms in the southwestern US. All farms were within an 8‐km radius and were sampled in August 2001, January 2002 and August 2002 (60 cows per farm per sampling; n = 720 total samples). Samples were cultured for E. coli O157:H7 and Salmonella and a portion of the recovered Salmonella isolates were examined for genetic relatedness using pulsed‐field gel electrophoresis (PFGE). Results:  Faecal shedding of E. coli O157:H7 and Salmonella varied considerably between farms and at the different sampling times. Large fluctuations in the percentage of positive animals were observed from summer to summer for both of these pathogens. Similarly, Salmonella serotype and serotype prevalence varied from farm to farm and within farm from one sampling time to another. Multiple Salmonella genotypes were detected for a number of serotypes and identical genotypes were found on different farms with one genotype of Salmonella Senftenberg identified on three of the four farms. Significance and Impact of the Study:  This study demonstrated the wide variability in pathogen shedding within and among dairy farms all located in a small geographical region and highlights the complexity of pathogen control at the farm level.

Aims: To examine factors affecting faecal shedding of the foodborne pathogens Escherichia coli O157:H7 and Salmonella in dairy cattle and evaluate antimicrobial susceptibility of these isolates. Methods: Faecal samples were obtained in replicate from lactating (LAC; n ¼ 60) and non-lactating (NLAC; n ¼ 60) Holstein cattle to determine influence of heat stress, parity, lactation status (LAC vs NLAC) and stage of lactation [£60 or >60 days in milk (DIM)] and cultured for E. coli O157:H7 and Salmonella. A portion of the recovered isolates were examined for antimicrobial susceptibility using the broth microdilution technique. Results: No effects of heat stress were observed. Lactating cows shed more (P < 0AE01) E. coli O157:H7 than NLAC cows (43% vs 32%, respectively). Multiparous LAC cows tended to shed more (P ¼ 0AE06) Salmonella than primiparous LAC cows (39% vs 27%, respectively). Parity did not influence (P > 0AE10) bacterial shedding in NLAC cows. Cows £60 DIM shed more (P < 0AE05) Salmonella than cows >60 DIM. Seventeen Salmonella serotypes were identified with the most prevalent being Senftenberg (18%), Newport (17%) and Anatum (15%). Seventy-nine of the Salmonella isolates were resistant to at least one of the seven antibiotics. Escherichia coli O157:H7 isolates were resistant to 11 different antibiotics with multiple resistance to nine or more antibiotics observed in five isolates. Significance and Impact of the Study: This study demonstrated differences in the shedding patterns of foodborne pathogens due to the stage of the milk production cycle and may help identify times when on-farm pathogen control would be the most effective.

Mammary epithelial cell (MEC) growth is reduced in continuously milked (CM) mammary glands, and administration of a mammogenic compound such as prostaglandin E(2) (PGE(2)) at parturition might improve MEC growth in CM tissue. The objectives were to 1) compare MEC turnover, ultrastructure, and gene expression in CM and involuting mammary tissue, and 2) evaluate the effects of CM and intramammary infusion of PGE(2) on early lactation MEC turnover, ultrastructure, mammary gene expression, milk yield, and composition. First- and second-lactation cows (n = 8) were used in a half-udder model, in which one-half was dry for 60 d (CTL) and the other was CM. Udder halves (n = 16) were assigned to a postpartum (PP) treatment of PGE(2) (+PGE(2); 875 mug/10 mL of medium-chain triglyceride oil) or no PGE(2) (-PGE(2)) treatment at parturition and at 72 h PP. Biopsies of CM and CTL quarters were obtained during milk stasis (MS) of the CTL half at 3 and 7 d after dry-off of the CTL half (3d-MS; 7d-MS) and postpartum (PP) at 2 and 4 d (2d-PP; 4d-PP). Milk yield was reduced (P < 0.01) in CM udder halves compared with CTL halves (13.2 vs. 22.1 kg/d), but reductions were less in second-lactation cows. The apoptotic index was greater (P < 0.05) in CTL glands than in CM glands (3d-MS, 0.52 vs. 0.11% and 7d-MS, 0.24 vs. 0.12, respectively). Proliferation of MEC was unchanged at 3d-MS, but was increased (P = 0.01) in CTL halves at 7d-MS compared with CM halves (3.10 vs. 0.93%). At 2d-PP, MEC proliferation was increased (P = 0.05) in CM halves compared with CTL halves (1.3 vs. 0.6%), but was unaffected by PGE(2) (P > 0.2). Apoptosis was elevated in early lactation regardless of treatment. Ultrastructure was unchanged by dry period length or PGE(2). In prepartum tissue, involution in CTL halves increased (P < 0.05) the expression of the proapoptotic genes Bcl-2-associated x protein (bax) and IGFBP5 and decreased (P < 0.05) alpha-lactalbumin expression compared with CM tissue. In PP mammary tissue, CTL halves expressed greater (P < 0.05) levels of ATP-binding cassette 1 (ABC1) and IGFBP5. Treatment with PGE(2) did not alter (P > 0.1) gene expression. The results confirm that CM reduced milk yield of cows with a mammary growth requirement. Reduced MEC turnover and milk yield were not alleviated by IMI of PGE(2), which indicates that peripartum PGE(2) concentrations in CM glands are not limiting mammary growth or milk synthesis.