Identification of elevated concentrations of estradiol in bovine uterine endometrium

We have investigated the estradiol content of bovine endometrium and related this to circulating plasma estradiol content. In 9 heifers, mean ± S.E.M. plasma estradiol concentration was 0.64 ± 0.25 pg/ml while the mean ± S.E.M. endometrial estradiol content was 43.0 ± 14.7 pg/g tissue; there was a close relationship between plasma and tissue estradiol levels (R² = 0.81; P < 0.001). During culture of endometrial tissue there was a progressive transfer of estradiol from tissue to culture media but no change in total estradiol. Culture of endometrium from 4 heifers with 5 ng/ml testosterone for 72 h resulted in no increase in estradiol. Furthermore, immunohistochemistry revealed no aromatase protein in uterine endometrium. These results confirm high stored tissue concentrations of estradiol in bovine endometrium while providing no evidence for estradiol synthesis by this tissue. The mechanism(s) through which this sequestration of estradiol into uterine tissue occurs remains to be determined.     

The absorption of phosphate ions from the ovine reticulorumen

The reticulorumen is now recognised to be an important site of net absorption of phosphate ions from ruminal fluid containing phosphate concentrations appropriate to those found in normal farming practice. These rates of absorption were measured in vivo from solutions placed in the washed reticulorumen, isolated in situ, in conscious, trained sheep. Reducing the ruminal sodium concentration led to reduced absorption of phosphate, suggestive that phosphate and sodium fluxes across the apical wall of the ruminal epithelial cell are linked, as they are in the kidney. Increased absorption of short chain fatty acids led to enhanced absorption of phosphate ions. Conversely, inhibition of carbonic anhydrase activity, by the addition of 1 mM acetazolamide to the ruminal fluid, led to a reduction in phosphate absorption. An increase in the acidity of the ruminal fluid also increased the absorption of phosphate, as did an increase in the ruminal Ca(2+) concentration over the range 1-4 mmol per litre. It is suggested that these effects can be accounted for by a Na(+)/H(+) antiporter coupled with a phosphate/proton symporter in the apical membrane of the ruminal epithelial cell.     

Disappearance and passage of propionic acid from the rumen of the beef steer

Studies were conducted to define steady state pH, propionic acid concentrations and fluid turnover in the rumen of steers fed every 3 h and to determine whether rates of ruminal propionic acid disappearance were linearly related to their in situ production rates. Ruminally fistulated beef steers (326 +/- 11 kg; n = 3) were fed eight times daily a 54% mixed hay: 46% corn-based concentrate diet to meet maintenance energy requirements. Maximal acceptable variations from the mean at steady state conditions of pH, propionic acid concentrations and specific activity, and liquid flow (Cr marker concentrations) were defined as 4%, 20%, 30% and 10%, respectively, across 4 h of observation. In situ production of propionic acid, determined by pulse-continuous infusion of 1-14C-propionic acid, was 142 mmol/h (CV = 8.4%). The ruminal half-life of propionic acid was estimated to be 1.5 h and the liquid flow rate was 3.8 liters/h. Propionic acid production rates subsequently were elevated by continuous intraruminal infusion of buffered propionic acid. Irrespective of production (basal and infusion) rate, approximately 66% disappeared (i.e., presumed absorption) and 34% passed from the rumen to the lower tract in the liquid phase. Ruminal disappearance of propionate was linearly related with its production rate, and propionate concentrations and production rates also were linearly related. Liquid passage was linearly related to production rate, but there was insufficient evidence to conclude that fractional dilution rate or ruminal volume were similarly related. When ruminal production of propionic acid is elevated, the rumen has additional absolute capacity to remove propionate, but the fractional removal appears to be constant. The digestive tract distal to the rumen is likely an important site of propionate absorption in cattle when propionate production is high.     

Effect of hemolysis on nonesterified fatty acid and beta-hydroxybutyrate concentrations in bovine blood.

Nonesterified fatty acid (NEFA) and beta-hydroxybutyrate (BHB) assays are used for evaluating dairy herds for negative energy balance and subclinical ketosis, respectively. Hemolysis is a common artifact in samples submitted to diagnostic laboratories. The effect of hemolysis on NEFA and BHB in bovine serum was determined. Hemolysis was introduced into 26 serum samples by adding serial dilutions of a red cell hemolysate, prepared by repeated freeze-thawing of EDTA-anticoagulated bovine blood. NEFA, BHB, and degree of hemolysis (hemolytic index) were measured by an automated chemistry analyzer. Two endpoint assays that differed by inclusion of a sample blank were used for NEFA measurement. A kinetic enzymatic assay with 2 reagent sources was used for BHB measurement. The assessed methods yielded similar NEFA or BHB results in baseline, nonhemolyzed samples (median NEFA: 0.25 mEq/L, median BHB: 3 mg/dL, median hemolytic index: 8 units). NEFA results were adversely affected by hemolysis, with values increasing significantly with higher degrees of hemolysis. Median values increased above a critical medical decision limit (0.40 mEq/L) at a hemolytic index of 506 units (marked hemolysis). This increase was prevented by inclusion of a sample blank. Result interpretation was affected in individual animals when samples were moderately hemolyzed (median hemolytic index: 258 units). In contrast, BHB results were unaffected by hemolysis with either reagent source. Thus, assays for measuring NEFAs should include a sample blank and NEFA results should not be interpreted in moderately to markedly hemolyzed bovine samples, because result accuracy cannot be assured.     

In vitro effects of 2,4-dichlorophenoxy acetic acid (2,4-D) on bovine cells.

Bovine fetal muscle cells were exposed to culture media containing 2 mg and 20 mg per liter of 2,4-dichlorophenoxy acetic acid (2,4-D) for varying intervals to determine the in vitro response of mammalian cells to this compound. The concentrations of 2,4-D used were comparable to those used in spray programmes although the residues normally found in pasture are much lower since 2,4-D is rapidly degraded under field conditions. Untreated and treated cultures were analyzed for total cell count, mitotic index and the percentages of differentiating and degenerating cells. The response of cultures to treatment was similar irrespective of the concentrations of 2,4-D used although in higher concentrations there was an initial drop in mitotic index. Other changes noted in treated cultures included an increase in differentiating and degenerating cells compared to those in control. The mitotic cells in treated cultures exhibited unipolar and tripolar spindles and a variety of other abnormalities including malorientation of the mitotic apparatus in relation to the axis of the cell. Myoblasts in initial stages of myogenesis were noted to be in mitosis in treated cultures suggesting that 2,4-D may have a stimulatory effect on myoblasts which in normal myogenesis are in post mitotic stage.     

Portovenogram findings in cases of elevated bile acid concentrations following correction of portosystemic shunts.

The case records of 36 cats and dogs undergoing surgical correction of a single extrahepatic portosystemic shunt were reviewed. In 12 animals, the shunt was fully ligated during the first surgical procedure, while, in the remaining 24, the shunting vessel could only be partially ligated. Assessment of serum bile acid concentrations demonstrated complete shunt occlusion in 15 of these latter 24 animals (63 per cent) between one and six months postoperatively. Ten animals (28 per cent) had persistently high serum bile acid concentrations postoperatively. Portovenogram findings in these individuals revealed six that demonstrated shunting solely through the original vessel; In five of these, full shunt attenuation was achieved at second surgery. Further shunt manipulation was not possible in the sixth case due to extensive adhesion formation. In the remaining four animals with raised bile acid concentrations, the portovenogram demonstrated shunting through the original vessel as well as the development of multiple acquired shunts.     

Immunoglobulin concentrations and bovine enterovirus inhibitors in fetal bovine fluids.

Fluids from 53 bovine fetuses ranging in age from 90 to 240 days were examined for immunoglobulin G (IgG) and immunoglobulin M (IgM) and neutralizing activity to ten bovine viruses. Non-specific inhibitors to bovine enteroviruses were found in serum, allantoic, and amniotic fluids of most samples tested. In most cases, serum IgG were within normal values. Neither IgG nor IgM was detected in amniotic fluids, whereas 2 samples of allantoic fluid contained traces of IgG.     

Disappearance of spermatoza from the ejaculates of vasectomized dogs.

Ejaculates of bilaterally vasectomized dogs contained spermatozoa as long as 21 days after vasectomy, indicating that spermatozoa in the canine ejaculate originated from both the epididymides and the vasa deferentia, not from the epididymides alone.     

Metabolism of propionate and 1,2-propanediol absorbed from the washed reticulorumen of lactating cows

To investigate the metabolism of 1,2-propanediol (PPD) in lactating cows independently of normal rumen microbial metabolism, three ruminally cannulated lactating Holstein cows were subjected to three experimental infusion protocols under washed reticulo-ruminal conditions in a Latin square design. Reticulo-ruminal absorption rates were maintained for 420 min by continuous intraruminal infusion of VFA and PPD. With the control treatment, 1,246 +/- 39 mmol/ h of acetate and 213 +/- 5 mmol/h of butyrate were absorbed from the reticulorumen. With the propionate treatment, 1,148 +/- 39 mmo/h of acetate, 730 +/- 23 mmol/h of propionate and 196 +/- 5 mmol/h of butyrate were absorbed from the reticulorumen. With PPD treatment, 1,264 +/- 39 mmol/h of acetate, 220 +/- 5 mmol/h of butyrate and 721 +/- 17 mmol/h of PPD were absorbed from the reticulorumen. Glucose irreversible loss rate (ILR), as well as the relative enrichment of plasma lactate and alanine, were determined by primed continuous infusion of [U-13C]glucose in a jugular vein. Treatments did not affect (P > 0.10) the plasma concentrations of glucose (4.2 +/- 0.1 mmoVL), alanine (0.14 +/- 0.01 mmol/L), or insulin (80 +/- 25 pmol/L). The plasma concentration of lactate was higher (P < 0.05) with both propionate (0.84 +/- 5 mmol/L) and PPD treatment (0.81 +/- 5 mmol/ L) compared with the control treatment (0.29 +/- 0.5 mmol/L). The plasma concentration of pyruvate was higher (P < 0.05) with the propionate treatment (0.09 +/- 0.01 mmol/L) compared with the control treatment (0.03 +/- 0.01 mmol/L). The plasma concentration of 3-hydroxybutyrate was lower (P < 0.05) with the propionate treatment (0.15 +/- 0.03 mmol/L) compared with the control treatment (0.40 +/- 0.03). With the PPD treatment, the plasma concentrations of pyruvate and 3-hydroxybutyrate were in between the other treatments and tended (P < 0.10) to be different from both. The plasma concentration of PPD increased throughout the infusion period with the PPD treatment and reached a concentration of 4.9 +/- 0.6 mmol/L at 420 min. The ILR of glucose was not affected (P > 0.10) by treatments (441 +/- 35 mmol/h). The relative 13C enrichment of plasma lactate compared with that of glucose decreased (P < 0.05) with the PPD treatment compared with the control treatment (44 to 21 +/- 3%). It was concluded that PPD has a low rate of metabolism in cows without a normal functioning rumen, although about 10% of the absorbed PPD was metabolized into lactate.     

Splanchnic metabolism of volatile fatty acids absorbed from the washed reticulorumen of steers

Six steers fitted with a ruminal cannula and chronic indwelling catheters in the mesenteric artery, mesenteric vein, hepatic portal vein, hepatic vein, as well as in the right ruminal vein were used to study metabolism of VFA absorbed from buffers in the emptied and washed reticulorumen. [2-(13)C]Acetate was infused into a jugular vein to study portal-drained visceral (PDV) uptake of arterial acetate, hepatic unidirectional uptake of acetate, and whole-body irreversible loss rate (ILR). Isobutyrate was infused into the right ruminal vein to calibrate VFA fluxes measured in the portal vein. On sampling days, the rumen was emptied and incubated in sequence with a 0-buffer (bicarbonate buffer without VFA), a VFA-buffer plus continuous intraruminal infusion of VFA, and finally another 0-buffer. Ruminal VFA absorption was determined as VFA uptake from the VFA-buffer and metabolic effects determined as the difference between metabolite fluxes with VFA-buffer and 0-buffers. Steady absorption rates of VFA were maintained during VFA-buffer incubations (4 h; 592+/-16, 257+/-5, 127+/-2, 17+/-<1, 20+/-<1 mmol/h, respectively, of acetate, propionate, butyrate, isovalerate, and valerate). The portal flux of acetate corrected for PDV uptake of arterial acetate accounted for 105+/-3% of the acetate absorption from the rumen, and the net portal flux of propionate accounted for 91+/-2% of propionate absorption. Considerably less butyrate (27+/-3%) and valerate (30+/-3%) could be accounted for in the portal vein. The sum of portal VFA and 3-hydroxybutyrate as well as lactate represented 99+/-3% of total VFA acetyl units and 103+/-2% of VFA propionyl units. Estimates are maximum because no accounting was made for lactate derived from glycolysis in the PDV. The net splanchnic flux of VFA, lactate, 3-hydroxybutyrate, and glucose accounted for 64+/-2% of VFA acetyl units and 34+/-5% of VFA propionyl units. Results indicate that there is a low "first-pass" uptake of acetate and propionate in the ruminal epithelium of cattle, whereas butyrate and valerate are extensively metabolized, though seemingly not oxidized to carbon dioxide in the epithelium but repackaged into acetate, 3-hydroxybutyrate, and perhaps other metabolites. When PDV "second-pass" uptake of arterial nutrients is accounted for, PDV fluxes of VFA, lactate, and 3-hydroxybutyrate represent VFA production in the gastrointestinal tract and thereby VFA availability to the ruminant animal.     

气相色谱法测定发酵液中丙酸和乙酸的含量

丙酸菌发酵法生产丙酸时,发酵液中同时积累丙酸和乙酸,用气相色谱仪可将发酵液样品中丙酸、乙酸两组分良好分离,并快捷测定发酵液中丙酸和乙酸的含量。气相色谱操作条件为:色谱仪进样器和检测器温度均为200℃;柱室温度150℃;氮气(高纯氮)流速为2.96ml/min;进样量2μl,外标法定量。丙酸保留时间8.4min,乙酸保留时间3.6min。丙酸浓度在500～4000mg/100ml范围内与峰面积呈线性相关,R=0.99875;乙酸浓度在200～2000mg/100ml范围内与峰面积呈线性相关,R=0.99956;该方法是一种简便、快速、准确的测定发酵液中丙酸和乙酸的方法。     

Characterization studies on mycoplasmas isolated from bovine mastitis and the bovine respiratory tract.

Mycoplasmas isolated from bovine mastitis in California were classified into five distinct species. These included Mycoplasma bovis, M. bovigenitalium, M. alkalescens, M. canadenfe, and an unidentified strain, ST-6. Strains frequently recovered from the nose of young calves proved to be M. arginini, M. bovirhinis was recovered from the respiratory tract but was not a common finding.     

Growth hormone, insulin and glucose concentrations in bovine fetal and maternal plasmas at several stages of gestation

For cows on d 137 (n = 6), 180 (n = 8), 226 (n = 9) and 250 (n = 5) of gestation (Exp. 1), concentrations of insulin and glucose were two- to three-fold less (P less than .01) in fetal venous plasma than in uterine arterial plasma. Concentrations of growth hormone, conversely, were 10- to 20-fold greater (P less than .01) in fetal venous than in uterine arterial plasma. Concentrations of insulin and glucose in maternal and fetal plasmas and concentrations of growth hormone in maternal plasma did not vary with stage of gestation. Concentrations of growth hormone in fetal venous plasma, however, were greater on d 226 and 250 than on d 137 and 180. For cows (n = 6) on d 198 of gestation (Exp. 2), concentrations of insulin and glucose in maternal and fetal plasmas and of growth hormone in maternal plasma remained relatively constant in samples collected every 30 min for 3 h. In contrast, growth hormone concentrations in fetal venous plasma were highly variable and appeared to be episodic, with pulses of 10 to 60 ng/ml in amplitude. No significant correlations were found among concentrations of insulin, glucose and growth hormone in fetal venous plasma. When samples were collected every 15 min for 4 h from cows (n = 5) on d 198 of gestation (Exp. 3), episodes of growth hormone in fetal venous plasma were irregular in amplitude and frequency.(ABSTRACT TRUNCATED AT 250 WORDS)     

Bactericidal effects of ozone at nonspermicidal concentrations.

A study was conducted to assess the use of ozone (O3) to control pathogens or contaminants of concern to animal breeders and regulatory officials. In separate experiments, samples of fresh bovine semen and Pseudomonas aeruginosa, Escherichia coli, or Campylobacter fetus subsp. venerealis were diluted with antibiotic-free milk (10(6) sperm and 10(6) organisms/mL of diluted semen), exposed in the previous day to a constantly monitored level of 5, 10, 15, or 20 micrograms/mL of O3 for 3-5 min. After 10 min at 30 degrees C, sperm motility was assessed and the samples cooled to 5 degrees C. Two and 18 h after the beginning of cooling, aliquots of each semen sample were evaluated for motility and cultured for organisms. Reductions were observed in P. aeruginosa and E. coli colony counts of 2 logs, and in C. fetus of 5 logs, after exposure for 2 h to O3 at a concentration of 5 micrograms/mL that had a moderate effect on sperm motility (reduction of 20%). Fewer than 100 colonies, i.e., a 4 logs reduction of all bacteria, were counted after dilution with ozonized-treated milk at 20 micrograms/mL of O3. However, this concentration of O3 reduced sperm motility by 50% 10 min after dilution. The results of these experiments indicate that a concentration and exposure time to O3 can be selected to reduce P. aeruginosa, E. coli, and C. fetus in contaminated bull semen diluted with milk while having only minimal effects on sperm motility.     

饲料有机酸化剂中乙酸、柠檬酸的高效液相色谱检测分析

应用反相液相色谱法同时分析测定饲料有机酸化剂中乙酸和柠檬酸的化学含量,结果表明,酸化剂中乙酸变异系数不超过2%,柠檬酸变异系数不超过3%;乙酸和柠檬酸的回收率分别为98%～101%、92%～96%。该方法表现出较高的准确性和精确度。