Prolactin and colibacillus-induced fluid transport in ligated intestinal segments.

Two experiments, using the ligated intestinal segment technique, were conducted to determine whether the pituitary hormone prolactin (PRL) could reduce Escherichia coli-induced fluid loss into the small intestine of 2- to 3-week-old pigs. Inoculation of 10(6) to 10(8) enteropathogenic E coli organisms into ligated jejunal segments caused a significant accumulation of luminal fluid within 12 hours. In the first experiment, intraluminal inoculation with 0.5 mg of ovine PRL along with the bacteria did not have any effect on fluid accumulation. Systemic IV treatment of the animals with 1.0 mg of ovine PRL at 3-hour intervals, beginning either immediately after or 9 to 10 hours before intestinal ligation, did not significantly (P less than 0.05) reduce fluid accumulation as compared with control animals. In the second experiment, IM administration of 100 microgram of thyrotropin-releasing hormone (TRH) at 3-hour intervals, beginning 6 hours before intestinal ligation, significantly (P less than 0.05) increased circulating PRL concentrations, as measured by radioimmunoassay. However, TRH treatment did not reduce the accumulation of luminal fluid in E coli-inoculated segments.     

Effect of dietary energy, body condition and calf removal on pituitary gonadotropins, gonadotropin-releasing hormone (GnRH) and hypothalamic opioids in beef cows

Beef cows (n = 64) were slaughtered to evaluate effects of dietary energy and calf removal (CR) on hypothalamic and adenohypophysial endocrine characteristics. From d 190 of gestation until parturition, cows received maintenance (ME; n = 32) or low (LE; n = 32) energy diets (ME = 100%, LE = 70% NRC recommendations). After parturition, half (n = 16) of each prepartum diet group received low (LE; n = 32) or high (HE = 130% NRC; n = 32) energy diets. At 30 d postpartum, cows were slaughtered 0 or 48 hr after CR. Hypothalami [preoptic area (POA), hypothalamus (HYP), stalk-median eminence (SME)] and pituitaries were collected. Basal and K(+)-induced release of GnRH from SME, and pituitary luteinizing hormone (LH) and follicle stimulating hormone (FSH) did not differ among groups (P greater than .05). Hypophyseal LH was correlated (P less than .01) with body condition score (BCS) at parturition and slaughter (r = .36 and .47, respectively). Prepartum LE diet increased (P less than .05) met-enkephalin in POA compared to prepartum ME (.59 +/- .05 vs. .44 +/- .04 pmol/mg) regardless of postpartum diet or suckling status. Concentrations of beta-endorphin in combined HYP + POA were decreased (P less than .05) by 48 hr CR (15.1 +/- 1.1 vs. 18.1 +/- 0.7 fmol/mg).(ABSTRACT TRUNCATED AT 250 WORDS)     

Aspartic Proteinase Members Secreted by the Ruminant Placenta: Specificity of Three Radioimmunoassay Systems for the Measurement of Pregnancy-associated Glycoproteins

Pregnancy‐associated glycoproteins (PAGs) isolated from the placenta of various ruminant species are enzymatically inactive members of the aspartic proteinase family. The measurement of these proteins in the maternal blood can be a good indicator of the presence of a live embryo. As certain aspartic proteinases are present in biological fluids in physiological and pathological conditions at various concentrations, it was necessary to determine the specificity of three radioimmunoassay (RIA) systems currently used for the detection of PAG molecules. Commercially available members of the aspartic proteinase family like pepsinogen, pepsin, chymosin, rennet, cathepsin D and renin were tested in a wide concentration range (10 ng/ml – 1 mg/ml). Pepsinogen cross‐reacted in RIA 1, RIA 2 and RIA 3 over 1 mg/ml, 50 μg/ml and 500 μg/ml concentrations, respectively. In the presence of pepsin, cross‐reaction was observed in RIA 1, RIA 2 and RIA 3 over 1 mg/ml, 500 μg/ml and 1 mg/ml concentrations, respectively. Chymosin and rennet could cross‐react in RIA 2 and RIA 3, while renin and cathepsin D did not decrease the binding of the tracer to antisera more, than that of the minimal detection limit. As the plasma/serum concentrations of the examined aspartic proteinases reported in the literature were outside the concentration range where cross‐reaction was observed, it can be concluded that these RIA systems were specific for the detection of PAGs in biological fluids.     

Relationships among concentrations of four opioid neuropeptides and luteinizing hormone-releasing hormone in neural tissues of beef cows following early weaning

Acute changes associated with removal of the inhibition of estrus caused by suckling were examined in beef cows. Calves were weaned during the fifth week after parturition and cows were slaughtered at 0 (n = 8), 36 (n = 8) or 72 h (n = 8) after calf removal. Tissues of preoptic area (POA), hypothalamus (HYP), pituitary stalk-median eminence (SME) and pituitary neurointermediate lobe (NIL) were obtained for analyses of luteinizing hormone-releasing hormone (LHRH) and four opioid neuropeptides. In addition, one-half of each SME was superfused in vitro for measurement of basal and potassium-induced release of LHRH. The following opioid neuropeptides were quantified: methionine-enkephalin (Met-Enk), beta-endorphin (beta-EP), dynorphin-A, 1-17 (DYN-17) and dynorphin-A, 1-8 (DYN-8). All four opioid neuropeptides were most concentrated in the pituitary NIL. Luteinizing hormone-releasing hormone was most concentrated in the SME tissue, which also contained substantial concentrations of Met-Enk and beta-EP, but very little DYN-17 or DYN-8. In addition, weaning increased the weight of NIL between 0 and 36 h (P less than .05), and the concentrations of LHRH, Met-Enk, and DYN-17 in the combined POA + HYP (P less than .05) tissue between 36 and 72 h. No differences occurred among groups in SME content of LHRH or in vitro release of LHRH from the superfused SME. Although they were not affected by weaning, within-cow correlations among parameters revealed that: 1) concentrations of DYN-17 and DYN-8 were always positively correlated (P less than .05); 2) concentrations of LHRH were positively correlated with Met-Enk (P less than .01), beta-EP (P less than .05) and DYN-17 (P less than .05) in the combined POA + HYP tissue; 3) LHRH concentrations in SME tissue were negatively related to POA + HYP concentrations of Met-Enk (P less than .01) and beta-EP (P less than .05), but not of LHRH or DYN-17 and 4) in vitro release of LHRH from the pituitary SME was correlated with concentrations of DYN-8 in various tissues including the SME (P less than .01). In summary, bovine neural tissues differ widely in concentrations of the four opioid neuropeptides with NIL tissue having the greatest concentrations. Weaning calves at 36 and 72 h before slaughter caused parallel changes in LHRH, Met-Enk and DYN-17 in preoptic and hypothalamic tissues.(ABSTRACT TRUNCATED AT 400 WORDS)     

Reduction in the mRNA expression of sVEGFR1 and sVEGFR2 is associated with the selection of dominant follicle in cows

The vascular endothelial growth factor (VEGF) is essential for follicular development by promoting follicular angiogenesis, as well as for the proliferation and survival of granulosa cells. The biological effects of VEGF are regulated by two membrane receptors, VEGFR1 and VEGFR2, and two soluble receptors, sVEGFR1 and sVEGFR2, which play an antagonistic role. Thus, the objective of this study was to identify the mRNA expression pattern of total VEGF, VEGFR1, VEGFR2, sVEGFR1 and sVEGFR2 in bovine preselected follicles (PRF) and post‐selected follicles (POF). The mRNA expression of these five genes in both granulosa cells (GC) and theca cells (TC) was compared between follicles classified as PRF and POF based on their diameter and on their ratio of estradiol/progesterone (E2/P4). Results showed a lower expression of mRNA of sVEGFR1 and sVEGFR2 in POF than in PRF (p < .05). Regarding the mRNA expression of total VEGF, VEGFR1 and VEGFR2, there was no difference between POF and PRF follicles (p > .05). Our results showed that the mRNA expression of VEGFR2 and sVEGFR1 was more abundant than the expression of VEGFR1 and sVEGFR2, while GC was the main source of mRNA for total VEGF. On the other hand, TC was the follicular compartment where the receptors were most expressed. Our results suggest that non‐dominant follicles maintain a greater concentration of the mRNA expression of both membrane and soluble VEGF receptors. On the other hand, follicular dominance is related to a reduction in the mRNA expression of sVEGFR1 and sVEGFR2, which may favour VEGF binding with VEGFR2 and, hence, improve the follicular health and development.     

Quantification of naloxone binding sites in brains from suckled beef cows during postpartum anestrus and resumption of estrous cycles

Thirty beef cows, approximately 3 yr of age, were randomly assigned to be slaughtered on d 7, 14, 28, 42 or 56 postpartum. Each cow suckled one calf until slaughter. Data from cows slaughtered on d 42 and 56 were pooled and further classified as anestrous or cyclic based on the presence of a corpus luteum and elevated serum concentrations of progesterone at slaughter. Specific binding of [3H]naloxone (3H-NAL) to homogenates of tissue from hypothalamus (HYP), preoptic area (POA) and basal forebrain (BF) was assessed using multiple-point Scatchard analyses. Nonspecific binding was estimated in the presence of 10(-6) M naloxone. Separation of bound from free 3H-NAL was achieved by centrifugation at 20,000 X g. Concentration (fmol/mg original tissue wet wt) of 3H-NAL binding sites in POA tissue was higher (P less than .05) on d 28 postpartum in anestrous cows than in cyclic cows on d 42 + 56 postpartum (2.58 +/- .32 vs 1.58 +/- .10). When all anestrous cows were compared with cyclic cows, concentrations of 3H-NAL binding sites in POA tissues and in BF tissue were higher (P less than .05) in anestrous cows (anestrous POA, 2.12 +/- .17, cyclic POA, 1.58 +/- .10; anestrous BF, 2.94 +/- .41, cyclic BF, 2.19 +/- .16). Compared across brain regions for all cows, the concentration of specific binding sites for 3H-NAL was greater (P less than .01) in BF (2.5 +/- .2) than in POA (1.9 +/- .1) and greater (P less than .01) in POA than in HYP (1.5 +/- .1).(ABSTRACT TRUNCATED AT 250 WORDS)     

In vitro Development of Buffalo Oocytes in Media-containing Fluids from Different Size Class Follicles

Studies were conducted to investigate the effect of supplementation of fluid from different sized class [small (SFF, < 3 mm), medium (MFF, 3-8 mm) and large (LFF, > 8 mm)] of normal and cystic (CFF) ovarian follicles in oocyte culture media on oocyte maturation rate and embryo development in vitro and to test the efficacy of follicular fluid (FF) from different size classes as a whole oocyte maturation medium. Results suggested that FF were capable of developing buffalo oocytes to embryonic stage in vitro although its efficacy was lower than that of serum. Regardless of high maturation rates after in vitro maturation (IVM) in media containing FF or IVM in whole FF, low blastocyst rates were obtained after in vitro fertilization (IVF) and culture of embryos. Follicular fluid from small follicles had significantly (p < 0.05) higher potential of developing buffalo oocytes to embryonic stage in vitro than that from medium and large follicles. Cystic FF was not capable of supporting development of buffalo oocytes in vitro.     

Effect of Semen Extender and Density Gradient Centrifugation on the Motility and Fertility of Turkey Spermatozoa

In the absence of commercially viable methods for cryopreserving turkey spermatozoa, new processing methods are required to extend the functional life of stored turkey spermatozoa for artificial insemination. The present study evaluates the efficacy of a new extender (Turkey Semen Extend) and investigates the use of density gradient centrifugation in processing turkey spermatozoa for artificial insemination. The new extender is compared with two commercially available turkey semen extenders, Beltsville Poultry Semen Extender and Ovodyl. Turkey spermatozoa in Turkey Semen Extend were still motile 20 h after collection, representing a considerable improvement over the other semen extenders (40%, 0% and 8% for Turkey Semen Extend, Beltsville Poultry Semen Extender and Ovodyl, respectively). A field trial on a commercial turkey farm showed improved fertilization rates following insemination of turkey hens with semen extended in Turkey Semen Extend (89.7%) compared with Beltsville Poultry Semen Extender (86.9%). This difference is statistically significant (p < 0.05). Processing on a density gradient, optimized for turkey spermatozoa, also increased sperm survival (50% gradient-prepared spermatozoa still motile after 18 h compared with <10% non-processed spermatozoa). Preliminary studies indicate that gradient preparation of spermatozoa may aid survival during cryopreservation.