Early lactation in dairy cattle is a period of severe negative energy balance (NEB) characterized by reduced blood glucose and insulin concentrations and elevated blood GH concentrations. The liver is refractory to GH during NEB and this uncoupling of the GH-IGF axis results in diminished plasma concentrations of IGF-I. Our objectives were to examine the effects of insulin administration during the immediate postpartum period on plasma IGF-I and GH concentrations and to examine the hepatic expression of total GH receptors (all GH receptor transcripts), GH receptor 1A (GHR 1A) and IGF-I. In addition, we examined adipose tissue for total GH receptor and IGF-I mRNA levels to establish the effects of chronic hyperinsulinemia on an insulin-responsive peripheral tissue. Holstein cows (n=14) were subjected to either a hyperinsulinemic-euglycemic clamp (insulin; INS) or saline infusion (control; CTL) for 96 h starting on day 10 postpartum. Insulin was infused i.v. (1 µg/kg body weight per h), blood samples were collected hourly, and euglycemia was maintained by infusion of glucose. Insulin concentrations during the infusions were increased 8-fold in INS compared with CTL cows (2·33 0·14 vs 0·27 0·14 ng/ml (S.E.M.); P<0·001) while blood glucose concentrations were not different between treatments (45·3 2·2 vs 42·5 2·2 mg/dl; P>0·1). Plasma IGF-I increased continuously during the insulin infusion, and reached the highest concentrations at the end of the clamp, being almost 4-fold higher in INS compared with CTL cows (117 4 vs 30 4 ng/ml; P<0·001). Hepatic expression of GHR 1A and IGF-I mRNA was low in CTL cows, but was increased 3·6-fold (P<0·05) and 6·3-fold (P<0·001) respectively in INS cows. By contrast, in adipose tissue the changes in gene expression in response to insulin were reversed with decreases in both total GHR and IGF-I mRNA. The expressions of GHR 1A and IGF-I mRNA in liver tissue were correlated in INS (r=0·86; P<0·05), but not CTL cows (r=0·43; P>0·1). Insulin appears to be a key metabolic signal in coupling the GH-IGF axis, thus orchestrating a marked elevation in circulating IGF-I concentrations.

Cattle with Johne's disease can shed live Mycobacterium avium subsp. paratuberculosis (MAP) in their milk, and MAP can survive under simulated commercial pasteurization conditions. In several studies conducted in the United Kingdom and Canada, MAP DNA has been detected in retail pasteurized milk samples; however, in one study in the United Kingdom viable MAP was identified in commercially pasteurized milk. A double-blind study involving two laboratories was undertaken to evaluate retail pasteurized whole milk in the United States. Marshfield Clinic Laboratories used solid culture medium (Herrold's egg yolk agar slants with mycobactin J and amphotericin B, nalidixic acid, and vancomycin), and TREK Diagnostic Systems, Research and Development used liquid culture medium (ESP culture system). Cultures at both laboratories were eonfirmed by PCR. A total of 702 pints of retail whole milk were purchased in three of the top five milk-producing states (233 from California, 234 from Minnesota, and 235 from Wisconsin) over a 12-month period and were tested for the presence of viable MAP. The criteria used for identifying samples as positive for viable MAP were similar to those followed by most laboratories (positive culture with PCR confirmation). The combined data from the two laboratories revealed the presence of viable MAP in 2.8% of the retail whole milk pints tested. Although the number of samples containing viable MAP was similar among states (P > 0.05), there was a seasonal effect on the presence of viable MAP in retail milk (P = 0.05). More MAP-positive samples were identified during the third quarter of the year (July through September). Of the 22 brands of retail milk tested, 12 (55%) yielded at least one sample positive for viable MAP.

Forty Holstein heifers [body weight (BW) = 126 kg] were blocked by BW into groups of 4, and, within each block, heifers were randomly assigned to one of four treatments. Twenty heifers had ad libitum access to a diet formulated to produce a BW gain of 0.8 kg/d (control diet), and 20 heifers had ad libitum access to a diet formulated to produce a BW gain of 1.2 kg/d. (high diet). Half of the heifers fed each diet were injected daily with bovine somatotropin (bST; 25 micrograms/ kg of BW). The high diet increased daily BW gain as well as body condition score. Injection of bST also increased daily BW gain, but did not affect body condition score. The high diet reduced age at puberty by 58 d, but did not affect BW, withers height at puberty, or pelvic area at slaughter. Injection of bST had no effect on age at puberty, but increased BW, withers height at puberty, and pelvic area at slaughter. The high diet did not affect mammary parenchymal DNA, RNA, or the ratio of RNA to DNA. The injection of bST increased mammary parenchymal DNA, RNA, and the ratio of RNA to DNA. The high diet was more cost effective for rearing dairy heifers from 120 d of age to potential breeding size (> or = 363 kg of BW and postpubertal) than was the control diet. In conclusion, the high protein, high energy diet increased growth rate without detrimental effects on mammary development. Injection of bST increased BW, skeletal size, and mammary development.

To determine effects of rapid prepubertal growth on first-lactation milk production, Holstein heifers were randomly assigned to one of three treatments. Thirty-five heifers were fed a standard diet to meet NRC recommendations and produce 0.8 kg of body weight (BW) gain/d (standard). Thirty-five heifers were fed a diet with higher energy (2.8 Mcal of metabolizable energy/kg) and protein (19.7% crude protein; high diet) to produce 1.2 kg of BW gain/d (high). Thirty-five heifers were fed the high diet and injected daily with bovine somatotropin (bST) (25 microg/kg of BW; high-bST). Diets were fed and bST was injected from 135 kg of BW until pregnancy was confirmed. Heifers were inseminated after BW exceeded 363 kg. Pregnant heifers were commingled and fed similar diets through gestation, parturition, and lactation. High and high-bST heifers had greater prebreeding average BW gains than standard heifers. Conversely, standard heifers had a greater average BW gain during gestation than high and high-bST heifers. High and high-bST heifers were approximately 90 d younger than standard heifers at first insemination and first parturition. Postpartum BW, body condition scores, and withers heights at parturition, and calving ease scores were not different among treatments. Standard heifers produced 14% more milk than high heifers but not more than high-bST heifers. The high-protein, high-energy diet decreased age at first parturition and first-lactation milk production, but did not affect reproduction. Injection of bST during the prepubertal growth period combined with the high diet decreased age at first parturition without reducing milk production.

Mycobacterium avium ssp. paratuberculosis (MAP) is the etiologic agent of Johne's disease in cattle. The disease causes diarrhea, reduced milk production, poor reproductivity, emaciation, and eventually death. Culture on Herrold's egg yolk agar is considered to be the definitive test for diagnosis of Johne's in cattle. This method has moderate sensitivity (30 to 50%) and is 100% specific; however, it can take up to 16 wk due to the slow growth of MAP. Currently, serum ELISA is used to screen herds for Johne's disease, but positive tests must be confirmed culturally or by PCR. The current research sought to evaluate an in-house direct fecal PCR procedure and directly compare it to ELISA using culture as the gold standard. Serum and fecal samples were collected from cows (n = 250) with unknown Johne's status. Fecal samples were processed for culture on Herrold's egg yolk agar and direct PCR. Serum samples were tested using the Parachek serum ELISA. Overall, 67/250 [26.8%, 95% confidence interval (CI) 21.4 to 32.8] animals were culturally confirmed to be shedding MAP. The PCR and ELISA detected 74/250 (29.6%, 95% CI 24 to 35.7) and 25/250 (10%, 95% CI 6.6 to 14.4), respectively. Culture and PCR were able to detect more positive animals than ELISA. Overall, direct fecal PCR was 70.2% sensitive and 85.3% specific when using culture as the gold standard. The ELISA method was 31.3% sensitive and 97.8% specific. When culture reported <10 cfu, the sensitivity and specificity of PCR and ELISA were 57.1 and 85.3%, and 4.8 and 97.8%, respectively. When culture reported 10 to <40 cfu, the sensitivity of PCR and ELISA were 75 and 50%, respectively. When culture reported > or =40 cfu, the sensitivity of PCR and ELISA were 100 and 88.2%, respectively. Specificity could not be calculated at these levels because there were no negative samples. The direct PCR outperformed the ELISA in detecting animals potentially infected with MAP and was not significantly different when compared with culture. The direct fecal PCR method described here provides faster results than traditional culture and is more sensitive than ELISA at detecting animals suspected of Johne's disease. These data support the use of PCR as an alternative method for screening herds for prevalence and diagnosis of Johne's disease.