Poor maternal nutrition during gestation in sheep reduces circulating concentrations of insulin-like growth factor-I and insulin-like growth factor binding protein-3 in offspring.

To determine if poor maternal nutrition alters growth, body composition, circulating growth factors, and expression of genes involved in the development of muscle and adipose of offspring, 24 Dorset and Shropshire ewes were fed either 100% (control fed), 60% (restricted fed), or 126% (over fed) of National Research Council requirements. Diets began at day 116 ± 6 of gestation until parturition. At parturition, 1 lamb from each control fed (CON), restricted fed (RES), and over fed (OVER) ewe was necropsied within 24 h of birth (1 d; n = 3/treatment) or reared on a control diet for 3 mo (CON = 5, RES = 5, and OVER = 3/treatment) and then euthanized. Body weights and blood samples were collected from lambs from 1 d to 3 mo. Organ weights, back fat thickness, loin eye area, and tissue samples (quadriceps, adipose, and liver) were collected at 1 d and 3 mo of age. The RES lambs weighed 16% less than CON (P = 0.01) between 1 d and 3 mo of age. In RES, there was a tendency for reduced heart girth at 1 d and 3 mo (P < 0.07) and back fat was reduced 36% at 3 mo (P = 0.03). Heart weight was 30% greater in OVER at 1 d when compared with RES lambs (P = 0.02). Serum IGF-I and IGFBP-3 were reduced in RES and OVER lambs (P < 0.05). Leptin tended to be greater in OVER lambs compared with CON at 1 d and 3 mo (P ≤ 0.08). Triiodothyronine was reduced in RES at 1 d (P = 0.05) and triglycerides tended to be greater in OVER at 3 mo (P = 0.07). In liver, there was a tendency for increased expression of IGF-I in OVER (P = 0.06) and decreased IGFBP-3 in RES (P = 0.09) compared with CON lambs at 1 d. In adipose tissue, adiponectin expression was decreased in RES (P = 0.05) at 3 mo. At 1 d of age, muscle expression of IGF-I tended to increase in RES (P = 0.06). In conclusion, poor maternal nutrition during gestation reduced growth rate in offspring which may be because of reduced circulating IGF-I and IGFBP-3 and decreased expression of IGFBP-3 in the liver.

Short communication: Expression of T-box 2 and 3 in the bovine mammary gland.

To increase our understanding of the mechanisms by which growth hormone (GH) and insulin-like growth factor (IGF)-I influence bovine mammary gland development, the potential roles of T-box2 (TBX2) and T-box3 (TBX3) were investigated. Although no information regarding expression of either transcription factor in the bovine mammary gland exists, it is known that TBX3 and its closely related family member, TBX2, are required for mammary gland development in humans and mice. Additionally, TBX3 mutations in humans and mice lead to ulnar mammary syndrome. Evidence is present in bone that TBX3 is required for proliferation and its expression is regulated by GH, an important regulator of mammary gland development and milk production. We hypothesized that TBX2 and TBX3 are expressed in the bovine mammary gland and that GH, IGF-I, or both increase TBX2 and TBX3 expression in bovine mammary epithelial cells (MEC). Bovine mammary gland tissue, MAC-T cells, primary MEC, and fibroblasts were obtained and TBX2 and TBX3 expression was determined by real-time reverse transcription PCR. In addition, TBX2 and TBX3 expression was examined in cells treated with 100 or 500 ng/mL of GH or 100 or 200 ng/mL of IGF-I for 24 or 48 h. Both TBX2 and TBX3 were expressed in bovine mammary tissue. Surprisingly, expression of TBX2 was only detected in mammary fibroblast cells, whereas TBX3 was expressed in all 3 cell types. Growth hormone did not alter TBX3 expression in MAC-T cells or MEC. However, IGF-I increased TBX3 expression in MAC-T, but not in primary MEC. We did not observe a change in TBX2 or TBX3 expression in fibroblasts treated with GH and IGF. Therefore, we concluded that (1) TBX2 and TBX3 are expressed in bovine mammary gland, (2) their expression is cell-type specific, and (3) IGF-I stimulates TBX3 expression in MAC-T cells.

Caprylic acid reduces Salmonella Enteritidis populations in various segments of digestive tract and internal organs of 3- and 6-week-old broiler chickens, therapeutically.

We investigated the efficacy of feed supplemented with caprylic acid (CA), a natural, 8-carbon fatty acid for reducing Salmonella enterica serovar Enteritidis colonization in commercial broiler chickens. In separate 3- and 6-wk trials, 1-d-old straight-run broiler chicks (n = 70 birds/trial) were assigned to a control group (challenged with Salmonella Enteritidis, no CA) and 2 replicates of 0.7 and 1% CA (n = 14 birds/group). Water and feed were provided ad libitum. On d 1, birds were tested for any inherent Salmonella (n = 2 birds/group). For the 3-wk trial, on d 5, birds were challenged with 8 log(10) cfu of Salmonella Enteritidis of a 4-strain mixture by crop gavage, and after 5 d postchallenge, birds (n = 2 birds/group) were euthanized to ensure Salmonella Enteritidis colonization. Caprylic acid was supplemented the last 5 d before tissue collection (n = 10 birds/group). For the 6-wk trial, on d 25, birds were challenged and confirmed for Salmonella Enteritidis colonization. The birds (n = 10 birds/group) were euthanized for tissue samples after CA supplementation for the last 5 d. Caprylic acid at 0.7 or 1% decreased Salmonella Enteritidis populations in cecum, small intestine, cloaca, liver, and spleen in both 3- and 6-wk trials. Body weight of birds did not differ between the groups (P ≥ 0.05). Further, to elucidate a potential antibacterial mechanism of action of CA, we investigated if CA could reduce Salmonella Enteritidis invasion of an avian epithelial cell line and expression of invasion genes hilA and hilD. The cell invasion study revealed that CA reduced invasive abilities of all Salmonella Enteritidis strains by ~80% (P < 0.05). Gene expression studies indicated that CA downregulated (P < 0.001) Salmonella invasion genes hilA and hilD. These results suggest that supplementation of CA through feed could reduce Salmonella Enteritidis colonization in broiler chicken and potentially reduces the pathogen's ability to invade intestinal epithelial cells by downregulating key invasion genes, hilA and hilD.

Evidence for a potential role of neuropeptide Y in ovine corpus luteum function.

Neuropeptide Y (NPY) is a neurohormone that is typically associated with food intake, but it has also been reported to affect the production of progesterone from luteal tissue in vitro. However, NPY has not been previously immunolocalized in the ovine ovary or in the corpus luteum (CL) of any species, and the effects of this neurohormone on luteal function in vivo are not known. Thus, we performed fluorescent immunohistochemistry (IHC) to localize NPY in the ovine ovary and used avidin-biotin immunocytochemistry (ICC) to further define the intracellular localization within follicles and the CL. We then infused NPY directly into the arterial supply of the autotransplanted ovaries of sheep to determine the in vivo effect of exogenous NPY on ovarian blood flow and on the luteal secretion rate of progesterone and oxytocin. Immunohistochemistry revealed that the NPY antigen was localized to cells within the follicles and CL, in the nerve fibers of the ovarian stroma, and in the vessels of the ovarian hilus. In the follicle, the NPY antigen was localized to nerves and vessels within the theca interna layer, and strong staining was observed in the granulosal cells of antral follicles. In the CL, NPY was localized in large luteal cells and in the vascular pericytes and/or endothelial cells of blood vessels, found dispersed throughout the gland and within the luteal capsule. In vivo incremental infusions of NPY at 1, 10, 100, and 1,000 ng/min, each for a 30-min period, into the arterial supply of the transplanted ovary of sheep bearing a CL 11 d of age increased (P< or =0.05) ovarian blood flow. The intra-arterial infusions of NPY also increased (P< or =0.05) in a dose-dependent manner the secretion rate of oxytocin, which was positively correlated (P< or =0.05) with the observed increase in ovarian blood flow. The infusions of NPY had a minimal effect on the secretion rate of progesterone, and similar intra-arterial infusions of NPY into sheep with ovarian transplants bearing a CL over 30 d of age had no significant effect on ovarian blood flow or on the secretion rate of progesterone. These results suggest that NPY acts on the luteal vascular system and the large luteal cells to rapidly stimulate blood flow and the secretion of oxytocin, respectively, which collectively implies a putative role for NPY during the process of luteolysis when increasing amounts of oxytocin are secreted from the ovine CL in response to uterine pulses of prostaglandin F2alpha.

Energy status, lipid-soluble vitamins, and acute phase proteins in periparturient Holstein and Jersey dairy cows with or without subclinical mastitis.

In a retrospective, case-controlled, observational study, associations among indices of negative energy balance, plasma lipid and lipid-soluble vitamin concentrations, plasma acute phase protein status, and occurrence of a new subclinical intramammary infection (IMI) during the periparturient period were determined. Cows were paired based on breed and expected parturition date (EPD) and monitored from the cessation of lactation through wk 8 of the subsequent lactation. A cow was identified as developing a new IMI if the intramammary pathogen isolated postpartum differed from that isolated in wk -9 (relative to EPD). Mean body condition score (BCS) of cows at wk -9 was 3.71 +/- 0.12. Fifteen Holstein and 15 Jersey dairy cows met the study selection criteria. Cows with a new IMI had greater body condition score, body weight, and body weight loss compared with cows that did not develop a new IMI. Prepartum plasma concentrations of beta-carotene were greater for Jersey cows with a new IMI compared with Jersey cows without a new IMI and Holstein cows, regardless of IMI status. However, there was a significant delay in recovery of plasma concentrations of beta-carotene postpartum for Jersey cows with a new IMI compared with Jersey cows without a new IMI. Plasma alpha-tocopherol, albumin, and retinol binding protein concentrations were greater during the periparturient period for cows without a new IMI. Plasma haptoglobin was increased at wk 1 postpartum for cows without a new IMI. Milk protein and lactose percentages and milk urea N were decreased and somatic cell counts were increased in cows identified with a new IMI compared with cows that did not develop a new IMI. Dairy cows with greater tissue energy stores prepartum and reduced plasma proteins, beta-carotene, and alpha-tocopherol had a greater risk for developing a new IMI during the periparturient period.

Growth rate and changes of the somatotropic axis in beef cattle administered exogenous bovine somatotropin beginning at two hundred, two hundred fifty, and three hundred days of age.

To determine the effects of bovine somatotropin (bST) treatment beginning at 3 ages on the growth rate and components of the somatotropic axis, 40 beef cattle (200 +/- 21 d of age) were randomly assigned to 1 of 4 treatments (10 animals/treatment). Three of the treatment groups received bST (33 mug/kg of BW) daily beginning at 200, 250, or 300 d of age until all animals reached 400 d of age; the fourth group served as controls (0 bST). Animals were housed in pens (5 animals per pen; 2 pens per treatment) and fed a diet formulated for an ADG of 1.2 kg/d. Feed intake (per pen) was measured daily, and BW was determined weekly. Blood samples (10 mL) and ultrasound measurements were collected at 200, 250, 300, 350, and 400 d of age. Serum concentrations of ST and IGF-I were determined by RIA and IGFBP-2 and -3 by ligand blot procedures. Overall, cattle gained 284.0 +/- 14.7 kg of BW with a treatment x week interaction (P < 0.01), such that during the treatment period ADG was 11.6, 8.7, and 15.8% greater (P < 0.05) in cattle treated with bST beginning at 200, 250, and 300 d, respectively, relative to controls during the same time frame. Average DMI was 13.6% less (P < 0.05) in bST-treated cattle than in controls. Increases in ADG coupled with a reduction in DMI resulted in 11.7, 14.0, and 26.4% increases (P < 0.01) in the efficiency of gain (G:F) in bST-treated cattle beginning at 200, 250, and 300 d of age, respectively, compared with contemporary controls. Backfat thickness increased (P < 0.05) over time, but the magnitude of the increase was less in the bST-treated cattle (treatment x week interaction; P < 0.05). Area of the LM increased (P < 0.05) over time but was similar across treatment groups. Serum concentrations of ST, IGF-I, and IGFBP-3 increased (P < 0.05), whereas IGFBP-2 decreased (P < 0.05) over time. The changes in the components of somatotropic axis were more pronounced in bST-treated cattle compared with controls, with the greatest magnitude of response in animals that began bST treatment at 300 d of age. In conclusion, the exogenous bST-induced growth response was greater in animals that began to receive bST administration at 300 d of age and received it for a shorter period (100 d) compared with animals that received bST beginning at 200 or 250 d of age.

The feeding value of extruded corn grain in a corn silage-based ration for high-producing Holstein cows and heifers during mid lactation.

The objective of this study was to investigate the feeding value of extruded corn in a corn silage-based ration for high-producing Holstein cows during mid lactation. Sixteen multiparous and 8 primiparous Holstein cows (106 +/- 49.7 d in milk; 43.7 +/- 5.27 kg of milk/d) were paired based on parity, days in milk, milk production, and body condition score and assigned to 1 of 2 dietary treatments in a randomized block design for 10 wk including a 2-wk adaptation period. Cows were fed a total mixed ration and milked 3 times per day. Diets contained 44% forage (3:1; corn silage:grass silage), 44.7% grain, and either extruded corn (EXC) or finely ground corn (FGC) at 11.3% of ration dry matter. No significant differences were detected in dry matter intake, milk protein yields, fat-corrected milk yields, or body condition score between cows fed FGC and cows fed EXC. Multiparous cows fed EXC produced more milk during wk 3 through 8 with a reduced milk fat content compared with multiparous cows fed FGC. Milk protein content was greater for primiparous cows fed EXC during wk 5 through 8 compared with primiparous cows fed the FGC ration. The major effect of feeding 2.7 kg/d of EXC compared with FGC was an increase in milk production and a reduction in milk fat content for multiparous cows, and an increase in milk protein content for primiparous cows.

Antibacterial effect of caprylic acid and monocaprylin on major bacterial mastitis pathogens.

Bovine mastitis is the most significant economic drain on the worldwide dairy industry. Concerns regarding poor cure rates, emergence of bacterial resistance, and residues in milk necessitate development of alternative therapeutic approaches to antibiotics for treatment of mastitis. A variety of free fatty acids and their monoglycerides have been reported to exert antimicrobial activity against a wide range of microorganisms. The objective of our study was to examine the efficacy of caprylic acid, a short-chain fatty acid, and its monoglyceride, monocaprylin, to inactivate common mastitis pathogens, including Streptococcus agalactiae, Streptococcus dysgalactiae, Streptococcus uberis, Staphylococcus aureus, and Escherichia coli. Milk samples containing 50 mM or 100 mM caprylic acid, and 25 mM or 50 mM monocaprylin were inoculated separately with a 3-isolate mixture of each of the 5 pathogens, and incubated at 39 degrees C. Populations of surviving bacteria were determined at 0 min, 1 min, 6 h, 12 h, and 24 h of incubation. Both caprylic acid and monocaprylin reduced all 5 pathogens by >5.0 log cfu/mL after 6 h of incubation. Among the bacterial species tested, Strep. agalactiae, Strep. dysgalactiae, and Strep. uberis were most sensitive, and E. coli was most tolerant to caprylic acid and monocaprylin. Results of this study indicate that caprylic acid and monocaprylin should be evaluated as alternatives or adjuncts to antibiotics as intra-mammary infusion to treat bovine mastitis.

The ontogeny of the somatotropic axis in Hereford calves from birth to one year of age and its response to administration of exogenous bovine somatotropin.

Administration of exogenous bovine ST (bST) increases growth rate, feed efficiency, and carcass quality in beef cattle. The magnitude of response to bST in beef cattle is variable and related to the age of the animal. Our objective was to determine the response of the somatotropic axis, in particular IGF-I, IGFBP-2, and IGFBP-3, to bST treatment from birth to 1 yr of age. Blood samples were collected before and after a single injection of bST (500 mg) every 50 d from birth to 1 yr of age in male and female Hereford calves. Body weights and serum concentrations of ST, IGF-I, IGFBP-2, and IGFBP-3 were determined. At birth, serum concentrations of ST, IGF-I, and IGFBP-3 increased (P < 0.05) following bST treatment. From 50 to 350 d of age, average concentrations of ST and IGF-I were greater (P < 0.05) in males, whereas IGFBP-2 concentrations were greater (P < 0.05) in females. No gender differences in IGFBP-3 concentrations were observed. Following bST treatment, IGF-I increased (P < 0.05) from 50 to 350 d of age, IGFBP-2 decreased (P < 0.05) from 50 to 200 d of age, and IGFBP-3 increased (P < 0.05) at 250 d of age. At 250 d of age, baseline concentrations of IGFBP-2 decreased (P < 0.05). Due to the positive response of IGFBP-3 and decreased baseline IGFBP-2 at 250 d of age, we conclude that this is an age at which the somatotropic axis is most responsive to exogenous bST, and it therefore may be an appropriate age to begin bST treatment in beef calves to realize the positive influence of bST on BW gain, feed efficiency, and carcass composition.

The ontogeny of the somatotropic axis in male and female Hereford calves from birth to one year of age.

Insulin-like growth factor-binding proteins-2 and -3 may play a role in age-dependent growth response to bovine ST (bST) treatment in cattle; however, samples have been collected at infrequent intervals and at limited time points. Therefore, the objective of this experiment was to examine the ontogeny of components of the somatotropic axis in Hereford calves from birth to 1 yr of age at weekly intervals to determine whether there is a certain age or time frame when the somatotropic axis may change and/or potentially become more responsive to exogenous bST administration. Blood samples and body weight measurements were collected from eight male and eight female Hereford calves once per week from birth to 1 yr of age. Serum concentrations of ST, IGF-I, IGFBP-2, and IGFBP-3 were determined. Males began to grow faster than females at approximately 16 wk of age (P < 0.05). Average concentrations of ST, IGF-I, and IGFBP-3 were greater in males than females (P < 0.01). Average concentrations of IGFBP-2 were greater in females than in males (P = 0.05). Concentrations of ST decrease with age (P < 0.01); however, the decrease occurred earlier in female calves. Concentrations of IGF-I and IGFBP-3 increased in males and females (P < 0.01), and concentrations of IGF-I began to plateau at approximately the same time as growth rate differences were observed (16 wk of age). Following an initial increase (birth to approximately 16 wk of age), concentrations of IGFBP-3 remained constant until approximately 43 wk of age. Concentrations of IGFBP-2 increased to approximately 10 wk of age (P < 0.05), followed by a decrease, and then, similar to IGFBP-3, remained constant until 43 wk of age. Correlations between average daily gain, ST, IGF-I, IGFBP-2, and IGFBP-3 were determined. Average daily gain was negatively (P < 0.01) correlated with ST and positively (P < 0.1) correlated with IGF-I. In females, ST was negatively (P < 0.01) correlated with IGF-I. Concentrations of ST were positively correlated (P < 0.01) with IGFBP-2 and IGFBP-3. Concentrations of IGFBP-2 were negatively correlated (P < 0.01) with IGF-I and positively correlated (P < 0.01) with IGFBP-3. In conclusion, serum concentrations of ST, IGF-I, IGFBP-2, and IGFBP-3 differed between male and fe-male calves. In addition, changes in components of the somatotropic axis occurred around the same time as males began to grow faster than females.

The influence of level of feeding on growth and serum insulin-like growth factor I and insulin-like growth factor-binding proteins in growing beef cattle supplemented with somatotropin.

The objective of this study was to determine the effects of level of feeding on growth, feed efficiency (gain:feed; G:F), body composition (BC), and serum concentrations of somatotropin (ST), IGF-I, and IGF-binding proteins (BP) in growing beef cattle supplemented with bovine (b) ST. In each of two consecutive years, 40 growing beef cattle were blocked by weight (average BW: yr 1 = 316 kg, yr 2 = 305 kg) and used in a 2 x 2 factorial arrangement with main effects of bST (0 or 33 microg x kg BW(-1) x d(-1)) and level of feed intake (ad libitum [AL] or 0.75 AL). Relative to uninjected cattle, treatment with bST increased ADG 9.6% (1.14 vs 1.25 kg/d; P < 0.05) and increased G:F 8.1% (12.3 vs 13.3 gain [g]:feed [kg]; P < 0.05), whereas ADG in AL animals was 39% greater than that in 0.75 AL animals (1.39 vs 1.00 kg/d; P < 0.05). There was a tendency (P = 0.10) for a bST x level of feeding interaction, such that the increase in ADG with bST was greater in AL cattle than in 0.75 AL cattle (10.6 vs 7.8%; P = 0.10). Serum concentrations of ST were greater in 0.75 AL cattle than in AL cattle (13.0 vs 8.6 ng/mL; P < 0.05) and in bST-treated cattle than in uninjected cattle (16.3 vs 5.2 ng/mL; P < 0.05). Due to a bST x level of feeding interaction (P < 0.01), the magnitude of the increase in serum ST to exogenous bST was greater (P < 0.01) in 0.75 AL cattle than in AL cattle. Relative to uninjected cattle, treatment with bST increased (P < 0.05) serum concentrations of IGF-I and IGFBP-3 and reduced (P < 0.05) concentrations of IGFBP-2. Similarly, AL cattle had greater (P < 0.05) serum concentrations of IGF-I and IGFBP-3 and reduced (P < 0.05) IGFBP-2 compared with 0.75 AL cattle. In summary, treatment with bST increased growth rate and G:F and stimulated serum IGF-I and IGFBP-3 while reducing IGFBP-2. Feeding at 0.75 ad libitum intake reduced the magnitude of response for each of these variables. Thus, limit-feeding may reduce the effect of exogenous bST on growth rate by blunting bST-induced increases in IGF-I and IGFBP-3 and bST-induced decreases in IGFBP-2.

Inactivation of Escherichia coli O157: H7 and Listeria monocytogenes by PR-26, a synthetic antibacterial peptide.

This study reports the antibacterial effect of PR-26, a synthetic peptide derived from the first 26 amino acid sequence of PR-39, an antimicrobial peptide isolated from porcine neutrophils. A three-strain mixture of Escherichia coli O157:H7 or Listeria monocytogenes of approximately 10(8) CFU was inoculated to a final concentration of 10(7) CFU/ml in 1% peptone water (pH 7.0), containing 50 or 75 microg/ml of PR-26, and incubated at 37 degrees C for 0, 6, 12, and 24 h; at 24 degrees C for 0, 12, 24, and 36 h; or at 10 or 4 degrees C for 0, 24, 72, and 120 h. Control samples included 1% peptone water inoculated with each pathogen mixture but containing no PR-26. The surviving population of each pathogen at each sampling time was determined by plating on tryptic soy agar with incubation at 37 degrees C for 24 h. At 37 degrees C, PR-26 decreased E. coli O157:H7 and L. monocytogenes populations by >5.0 log CFU/ml at 12 h, with complete inactivation at 24 h. At 24 degrees C, PR-26 reduced E. coli O157:H7 and L. monocytogenes by approximately 3.5, 4.0, and 4.5 log CFU/ml at the end of 12-, 24-, and 36-h incubations, respectively. At 4 and 10 degrees C, the inhibitory effect of PR-26 on E. coli O157:H7 and L. monocytogenes was significantly lower (P < 0.05) than that at 37 and 24 degrees C: a 2- to 3-log CFU/ml reduction was observed at 120-h incubation. Results indicate that PR-26 could potentially be used as an antimicrobial agent, but applications in appropriate foods need to be validated.

Effect of dietary α-tocopherol supplementation on color and lipid stability in pork.

Myoglobin and lipid oxidation are major causes of quality deterioration in fresh pork. A process to enhance color and lipid stability would prove valuable to the pork industry given the current trend of centralized packaging and distribution to retail markets. Our objective was to determine the effects of dietary α-tocopherol (α-Toc) supplementation on color and lipid stability in ground pork, and loin chops stored in modified atmosphere packaging (MAP). Yorkshire crossbred pigs (n=20) were randomized into two groups and fed diets containing 48 (CON) or 170 mg α-Toc acetate/kg feed (VIT-E) for 6 weeks before slaughter. Plasma α-Toc concentration was measured weekly. Post-slaughter, Boston butt shoulders were ground, formed into patties with or without 1.5% salt, and stored fresh at 4°C for 0, 2, 4, or 6 days, and frozen at -20°C for 45 or 90 days. Pork loin chops were packaged aerobically and stored at 4°C for 0, 2, 4 or 6 days, or in MAP at 4°C for 7, 10 or 13 days prior to Hunter L*,a*,b* and TBARS analyses. α-Toc concentration of longissimus dorsi, psoas major, biceps femoris, semimembranosus and semitendinosus muscles was determined. Plasma α-Toc was greater (P<0.05) in VIT-E animals compared with CON and α-Toc concentrations were greater (P<0.05) in all VIT-E muscles compared with CON. TBARS values of both fresh and salted patties were less in VIT-E than in CON meat following 6 days at 4°C; VIT-E TBARS of salted patties were less (P<0.05) after 45 days at -20°C compared with CON. α-Toc supplementation did not influence (P>0.05) color of aerobically packaged or MAP chops, or of fresh or salted pork patties. α-Toc supplementation reduced TBARS formation in fresh and salted pork but had no significant impact on color.

Influence of somatotropin and nutrition on bovine oocyte retrieval and in vitro development.

The objective of this study was to determine the effects of supplemental bovine somatotropin (bST) and limit feeding on follicular growth and oocyte competence in yearling beef heifers. Sixteen growing heifers (424+/-4 kg) were randomly assigned to 1 of 4 treatments in a 2 x 2 factorial arrangement, with main effects of bST (0 or 33 microg/kg BW/d) and feeding regimen (ad libitum or 0.75 ad libitum intake). Animals were treated for 100 d prior to follicular aspiration, and treatments continued for the 42-d period that follicles were aspirated. Follicles were observed ultrasonically then aspirated, and recovered oocytes were matured, fertilized and developed in vitro. The number of follicles observed ultrasonically was greater with bST treatment (P<0.01) but was unchanged by plane of nutrition. The number and quality of recovered oocytes were similar among treatments, as was the number of oocytes resulting in blastocyst formation.

Expression of the human alpha1,2-fucosyltransferase in transgenic pigs modifies the cell surface carbohydrate phenotype and confers resistance to human serum-mediated cytolysis.

Hyperacute rejection (HAR) is the first critical immunological hurdle that must be addressed in order to develop xenogeneic organs for human transplantation. In the area of cell-based xenotransplant therapies, natural antibodies (XNA) and complement have also been considered barriers to successful engraftment. Transgenic expression of human complement inhibitors in donor cells and organs has significantly prolonged the survival of xenografts. However, expression of complement inhibitors without eliminating xenogeneic natural antibody (XNA) reactivity may provide insufficient protection for clinical application. An approach designed to prevent XNA reactivity during HAR is the expression of human alpha1, 2-fucosyltransferase (H-transferase, HT). H-transferase expression modifies the cell surface carbohydrate phenotype of the xenogeneic cell, resulting in the expression of the universal donor O antigen and a concomitant reduction in the expression of the antigenic Galalpha1,3-Gal epitope. We have engineered various transgenic pig lines that express HT in different cells and tissues, including the vascular endothelium. We demonstrate that in two different HT transgenic lines containing two different HT promoter constructs, expression can be differentially regulated in a constitutive and cytokine-inducible manner. The transgenic expression of HT results in a significant reduction in the expression of the Galalpha1,3-Gal epitope, reduced XNA reactivity, and an increased resistance to human serum-mediated cytolysis. Transgenic pigs that express H-transferase promise to become key components for the development of xenogeneic cells and organs for human transplantation.

Methionine and somatotropin supplementation in growing beef cattle.

This objective of this study was to determine the effects of supplemental bovine somatotropin and supplemental ruminally protected methionine (Met) on ADG, feed efficiency (FE), body composition (BC), and serum concentrations of Met, ST, IGF-I, and progesterone in growing beef cattle. Two experiments were conducted. In one experiment, 24 growing beef cattle (6 steers and 18 heifers, 410 +/- 2 kg) were divided into two treatment groups, either controls or Met-treated animals. In a second experiment, 32 heifers and 8 steers (311 +/- 2 kg) were used in a completely randomized design experiment (four treatments; two pens/treatment). Treatments were arranged as a 2 x 2 factorial, with main effects of ST (0 or 33 microg/kg BW per day) and Met (0 or 6 g x animal(-1) x d(-1)). Feed efficiency and ADG were not affected by Met. However, ST increased FE and ADG (P < .05). Serum Met concentrations tended (P = .08) to increase with Met administration and remained unchanged by ST administration. Serum ST and IGF-I concentrations were increased (P < .05) by ST administration but unchanged by Met administration. The BC and heifer serum progesterone concentrations were similar across treatments, and no interactions (P > .10) were observed between ST and Met. In summary, ST improved BW gain and FE. Methionine was not the first-limiting AA in the diet fed.

Rapid estimation of spoilage bacterial load in aerobically stored meat by a quantitative polymerase chain reaction.

We report a quantitative PCR which utilizes primers from a conserved 23S rDNA sequence identified in nine different spoilage bacteria commonly present in meat. The PCR detected the spoilage bacteria by amplifying a specific 207 bp sequence from their chromosomal DNA. Quantification of PCR product by electrochemiluminescence revealed that the concentration of the amplified product was dependent on cycle number and the initial number of bacteria present in the sample. Statistical analysis of the results indicated a correlation coefficient of 0.94 (P < 0.001) between aerobic plate count and QPCR luminosity units.

The role of phosphoinositide-derived second messengers in oxytocin-stimulated prostaglandin F2 alpha release from endometrium of pigs.

The mechanism for prostaglandin (PG) F2 alpha release from pig endometrium after oxytocin (OT) treatment is unknown. OT may rapidly stimulate inositol (1,4,5)-trisphosphate (IP3) and diacylglycerol (DAG) formation, consistent with the concept of rapid activation of a second-messenger system. In support of this hypothesis, endometrial IP3 levels were increased (P < 0.05) within 0.5 min after treatment with 0.1 microM OT. In contrast, increased DAG formation was not detected after treatment with OT. However, similar to the stimulation of endometrial PGF2 alpha secretion observed after OT treatment (P < 0.001), PGF2 alpha release was increased (P < 0.01) after treatment with phorbol-12-myristate-13-acetate (PMA), which mimics DAG activation of protein kinase C. Further, stimulation of endometrial PGF2 alpha secretion did not result from cell death induced by PMA or OT because lactate dehydrogenase, a cytosolic marker of cellular integrity, did not leak into the medium after PMA or OT treatment. In contrast, 0.5% saponin (positive control for cell death and concomitant release of lactate dehydrogenase) increased PGF2 alpha secretion (P < 0.05) and lactate dehydrogenase release (P < 0.001). These results indicate that OT induces endometrial IP3 production in a rapid manner indicative of a second-messenger system. The finding that increased DAG was not also detected after OT treatment may reflect rapid metabolism or compartmentalized production of DAG involved in the second-messenger stimulation of phospholipase C. The high background of DAG used in the biosynthesis of cellular lipids would obscure the rather small spatially localized changes in DAG levels resulting from the activation of phospholipase C. The finding that DAG was present at approximately 10 to 20-fold higher levels than IP3 in resting cells was consistent with this conclusion.

A proposed role for oxytocin in regulation of endometrial prostaglandin F2 alpha secretion during luteolysis in swine.

Pulsatile secretion of endometrial prostaglandin (PG)F2 alpha is stimulated by oxytocin (OT) during late diestrus in domestic ruminants (i.e., cattle, sheep and goats) and results in corpus luteum (CL) regression leading to the onset of a new estrous cycle. Pulsatile PGF2 alpha release is also responsible for CL regression in swine, but the stimulus for its secretion from the uterine endometrium is not known. We propose that OT binds to specific OT receptors (OTR) on the endometrium to stimulate phosphoinositide (PI) hydrolysis, thereby activating the inositol trisphosphate (IP3)-diacylglycerol (DAG) second-messenger system to promote pulsatile PGF2 alpha secretion. Exogenous OT administered to cyclic gilts during late diestrus (days 10-16) decreased interestrous interval in three of four experiments. However, OT did not promote CL regression in hysterectomized gilts indicating that the effect of OT was uterine-dependent. Circulating concentrations of 13,14-dihydro-15-keto PGF2 alpha (the major stable metabolite of PGF2 alpha) were increased (p < 0.01) 10 min after i.v. injection of OT on days 14 and 16 in cyclic gilts and on days 10-16 in pregnant gilts, but the magnitude of the response to OT on all days in pregnant gilts was markedly reduced compared to the response in cyclic gilts on days 14 and 16. Mean density and Kd of OTR detected on endometrium of cyclic pigs 15 days post-estrus were 29.2 +/- 5.5 fmol/mg protein and 1.59 +/- 0.23 nM, respectively. Density of OTR was correlated with OT-stimulated PI hydrolysis (r = 0.83, p < 0.05) and PGF2 alpha secretion (r = 0.87, p < 0.10). Endometrial IP3 was increased within 30 seconds after OT treatment and preceded the increase in PGF2 alpha release stimulated by OT. Endometrial PI hydrolysis and PGF2 alpha secretion were similarly increased by AIF4-(phospholipase C activator), but not by cholera toxin (adenylyl cyclase activator). Although OT binding to OTR could be displaced by lysine-vasopressin and lysine-vasopressin stimulated PI hydrolysis, lysine-vasopressin did not stimulate PGF2 alpha release. Distinct receptors for OT and lysine-vasopressin on pig endometrium were confirmed by treatment with 100 nM OT + 100 nM lysine-vasopressin which stimulated PI hydrolysis more than 100-200 nM OT or lysine-vasopressin alone. These results support the hypothesis that OT stimulates phospholipase C to hydrolyze PI, yielding IP3 and DAG second-messengers which promote endometrial PGF2 alpha release during CL regression in pigs.

Modifications in growth and morphology of ovine jejunal and ruminal epithelia as affected by inert dietary substances.

Thirty-two lambs were used to study the effects of a nondegradable material in the alimentary tract on growth and morphology of ruminal and jejunal epithelia. Lambs were fed isoenergetic rations that differed only by addition of ground polyethylene. Eight of the lambs receiving each diet were given polyurethane cubes orally that were confined to the rumen. Lambs were slaughtered after 30 d; morphological and growth characteristics of the anterior-ventral and posterior-dorsal ruminal epithelia and jejunal epithelia were examined. Weights of the stomach complex were greater (P less than .05; 1.57 vs 1.48 kg) for lambs fed the polyethylene diet. Polyurethane cubes had no effect on weights of the stomach complex and small intestines. Deoxyribonucleic acid levels and both height and width of papillae were greater (P less than .05) in ruminal epithelial from lambs fed the diet with polyethylene than in those from lambs given the control diet. Protein and DNA levels in jejunal epithelia also were affected (P less than .05) by diet, averaging 8.2 mg/cm2 and 510 micrograms/cm2, respectively, for lambs fed the polyethylene diets vs 7.3 mg/cm2 and 417 micrograms/cm2 for lambs fed the control diet, respectively. Average villus height was greater (P less than .05) in jejunal epithelia of lambs fed the control diet than in jejunal epithelia of those given the polyethylene diet (553 vs 466 microns). Polyurethane cubes did not affect growth or morphological characteristics of either ruminal or jejunal epithelia.(ABSTRACT TRUNCATED AT 250 WORDS)