Consequences of different dietary energy sources during follicular development on subsequent fertility of cyclic gilts

The objective of the present study was to investigate the effects of dietary-induced insulin enhancement during the late luteal phase on subsequent fertility of gilts. Fifty-two littermate cyclic gilts were subjected to dietary treatments where two energy sources were tested: corn starch (T1) and soybean oil (T2). The experimental diets were supposed to provide similar amounts of dietary energy, but from different sources. Gilts were fed ad libitum, starting day 8 of the estrous cycle, until the next standing heat. Blood sampling was performed in a subgroup of 20 gilts on days 14 and 21 of the cycle for analyses of glucose and insulin, and after ovulation detection until 18 h after ovulation for progesterone. All gilts were slaughtered on day 28 of pregnancy and the reproductive tracts recovered for further analysis. T1 gilts showed higher postprandial insulin peak on days 14 and 21 and lower glucose levels 4 h after feeding on day 14 (P<0.05), however, there were no treatment effects on plasma progesterone concentrations. Dietary energy sources did not affect average daily feed intake, body weight and backfat on day 28 of pregnancy. Estrous cycle length, estrus duration and time of ovulation were not affected by previous nutritional treatments either. T1 gilts showed higher ovulation rates, number of embryos, embryo weight and placental weight (P<0.05). There were no treatment effects on pregnancy rate, embryo survival rate and volume of amniotic fluid. A positive correlation between progesterone concentration 18 h after ovulation and ovulation rate was observed (r=0.75; P<0.01). These results suggest that it is possible to manipulate dietary insulin response in cyclic gilts and, thus, improve reproductive efficiency when feeding starch as the main energy source during the late luteal and follicular phases of the cycle.     

Intrauterine inoculation of minipigs with Chlamydia trachomatis during diestrus establishes a longer lasting infection compared to vaginal inoculation during estrus

Advanced animal models, such as minipigs, are needed for the development of a globally requested human Chlamydia vaccine. Previous studies have shown that vaginal inoculation of sexually mature Göttingen minipigs with Chlamydia trachomatis resulted in an infection lasting only 3–5 days.The aim of this study was to evaluate the effect of targeting the upper porcine genital tract by transcervical and transabdominal intrauterine inoculation, compared to previously performed vaginal inoculation. Furthermore, we investigated the effect of the hormonal cycle, estrus vs. diestrus, on the establishment of a C. trachomatis infection in the minipig.Targeting the upper genital tract (transcervical inoculation) resulted in a longer lasting infection (at least 7 days) compared to vaginal inoculation (3–5 days). When comparing intrauterine inoculation during estrus and diestrus, inoculation during diestrus resulted in a longer lasting infection (at least 10 days) compared to estrus (3–5 days). Furthermore, we found a significant C. trachomatis specific IFN-γ response in pigs inoculated during estrus correlating with the accelerated clearance of infection in these pigs.These findings suggest that for implementation of an optimal model of C. trachomatis in minipigs, inoculation should bypass the cervix and preferable be performed during diestrus.     

A clinical approach to determine false positive findings of clinical endometritis by vaginoscopy by the use of uterine bacteriology and cytology in dairy cows

Clinical endometritis in dairy cows is defined as mucopurulent or purulent vulvar discharge 21 days or more after parturition. The diagnosis of clinical endometritis is commonly based on vaginal examination. Techniques to reduce the proportions of false negative findings have been described. This paper discusses a clinical approach to determine the proportion of false positive findings that might occur by vaginal inspection. The consequences of false positive findings in dairy practice are unnecessary or inadequate treatments. In research, incorrect diagnoses have an impact on the interpretation of studies on the diagnosis and treatment of clinical endometritis. The objective of the present study was to compare intrauterine bacteriology and endometrial cytology in cows diagnosed with clinical endometritis with findings obtained by vaginoscopy. Clinical endometritis was defined as mucopurulent or purulent vulvar discharge. On two commercial dairy farms, cows were examined 21 to 28 d postpartum. Uterine samples (n = 230) were collected from cows with clinical endometritis with the cytobrush technique to determine the proportion of polymorphonuclear neutrophils (PMN) and to culture smears for aerobic bacteria. Two threshold values for the proportion of PMN (5 and 18%) were chosen as possible indicators for an inflamed endometrium. Common uterine pathogens A. pyogenes and E. coli were found in 33.5 and 10.4% of the samples, respectively. With increasing vaginal discharge score, proportion of samples positive for A. pyogenes increased significantly. The proportion of cows exceeding the thresholds for PMN increased with vaginal discharge score and the presence of A. pyogenes.Considering only the presence of aerobic uterine pathogens and a proportion of PMN above the threshold values of 5 and 18% as indicative for endometritis, a proportion of 17.3 and 28.5%, respectively, of diagnoses by vaginoscopy were false positive.     

Comparison of PGF2α-induced luteolysis in early pregnant and estrogen-treated ‘pseudopregnant’ gilts

Conceptus estrogen clearly plays a major role in luteal maintenance in the pig; however, other conceptus-derived substances or conceptus-induced uterine secretory products appear to have a local luteotrophic/anti-luteolytic effect on the corpora lutea (CL) and likely may play a key role in maternal recognition of pregnancy in the pig. The objective of these studies was to compare PGF₂α-induced luteolysis in estrogen-treated ‘pseudopregnant’ gilts versus pregnant gilts during the period of maternal recognition of pregnancy. In Experiment 1, doses of PGF₂α ranging from 1 to 100 μg were administered via intraluteal silastic implants to pseudopregnant gilts to determine the dose necessary to cause functional (progesterone) and structural (weight) luteal regression similar to that observed during the natural estrous cycle. Luteal sensitivity to this minimally effective luteolytic dose of PGF₂α was then determined for both pseudopregnant and pregnant gilts in Experiment 2. Experiment 3 investigated whether Day 13 porcine conceptus tissue could directly prevent PGF₂α-induced luteolysis at the level of the CL. The minimally effective luteolytic dose of PGF₂α (100 μg) determined in the pseudopregnant pig caused a similar decline in progesterone concentration and weight of CL in pregnant gilts, suggesting that the susceptibility of CL of pregnant and pseudopregnant pigs to PGF₂α is similar. However, luteal weight was greater (P<0.05) for the pregnant gilts than for pseudopregnant gilts, suggesting that estrogen treatment alone cannot mimic the conceptus effects on CL growth and development. Experiment 3 demonstrated that lyophilized Day 13 conceptus tissue implanted directly into individual CL could partially inhibit PGF₂α-induced luteolysis, providing for the first time direct evidence that porcine conceptuses as early as Day 13 contain factors which can directly (i. e. at the level of the CL) prevent luteal regression.     

Optimum Culture Conditions for Production of Exfoliative Toxin by Staphylococcus hyicus

Optimum culture conditions for the production of exfoliative toxin by Staphylococcus hyicus (shET) were examined. High shET activity was obtained from the culture filtrate of HI and TY broth inoculated with S. hyicus. The pH in these two media ranged from 7 to 8.5 during bacterial culture, while the lowest pH in TS and BHI broth was less than 6. shET activity in the culture filtrate from TY broth inoculated with 10⁷ CFU of S. hyicus per ml was higher than that in TY broth inoculated with 10⁶ and 10⁸ CFU of bacteria per ml. When shET activity in the culture filtrate was measured under various shaking conditions, the culture filtrate shaken at 75 oscillations per min had the highest shET activity of the five shaking conditions. shET activity of the culture filtrate of TY broth to which protease inhibitor had been added was the same as that of TY broth without inhibitor. shET activity in a shaking culture in an Erlenmeyer flask was also the same as that in sac culture and that in shaking culture using a shaking (Sakaguchi) flask. shET activity in TY broth supplemented with 100 mM glucose was significantly lower than that in TY broth without glucose. Based on the above results, the optimum culture conditions for the production of shET were as follows: inoculation of 3 × 10⁹ CFU of S. hyicus strain P-1 into 300 ml of TY broth in a 2,000-ml Erlenmeyer flask, and incubation at 37 C with shaking at 75 oscillations per min. Then shET activity of the culture filtrate under appropriate culture conditions was measured after various incubation periods. shET activity was detected 6 hr after inoculation, reached the maximum (2⁵³ exfoliative unit/0.1 ml) at 16 hr and decreased between 20 and 48 hr. Thus, the optimum incubation period was determined to be 16 hr. Then the optimum concentration of ammonium sulfate for isolation of shET from the culture filtrate under appropriate culture conditions was examined. The greatest shET activity was obtained from the fraction salted out with 90% saturated ammonium sulfate. Thus, the optimum concentration of ammonium sulfate for the isolation of shET was determined to be 90% saturation.     

Comparative analysis of functions of cow and reindeer female reproductive organs: Progesterone content in vessels of reproductive organs

Progesterone content in blood from paired ovarian and uterine veins as well as from jugular veins of cows and reindeers was studied in the estrous cycle lutein phase and at the earlier stages of pregancy. In the both species, maximal progesterone concentration was detected in blood from vein of the ovary carrying corpus luteum (p < 0.001). In cows, a higher hormone concentration, as compared with jugular vein, has also been determined in vein of the uterus horn closest to ovary with corpus luteum (p < 0.01). In reindeers, blood from all studied vessels of reproductive organs had the progesterone concentration that was statistically significantly higher (p < 0.001) than that from jugular vein. In cows, progesterone concentration in blood from the ovarian vein was found to be higher when corpus luteum was located on the right ovary (p < 0.05) as compared with left-side corpus luteum location. No functional asymmetry of ovaries was revealed in reindeers. A possible role of mechanisms of the hormone local transport between ovary and uterus in adaptation of ruminants to reproduction under Nordic conditions is discussed.     

Effect of estradiol and progesterone on oviductal LH-receptors and LH-dependent relaxation of the porcine oviduct

We have previously shown that the porcine oviduct possesses immunoreactive and functional LH receptors and that LH causes relaxation of the oviduct, especially during the periovulatory stage of estrous cycle. The current studies were undertaken to investigate the effects of estradiol and progesterone on LH receptor protein and LH-stimulated motility of the oviduct in steroid-primed ovariectomized gilts. Twenty-one cross-bred gilts were ovariectomized at 6 m.o. of age. Four weeks later gilts received daily intramuscular injection of either 2 mL corn oil (control n = 4), estradiol benzoate (EB) 1.5 mg (n = 6), progesterone 50 mg (n = 5), or 1.5 mg EB plus 50 mg progesterone (n = 6) for 4 consecutive days. The gilts were slaughtered on Day 5 after the first injection of steroids or vehicle. Rings of isthmus and ampulla were collected from each oviduct and placed in a tissue chamber perfused with Kreb's solution for 60 min. The mechanical activity was recorded for 30 min after LH treatment. Immunoreactivity of LHR in the Fallopian tube sections were detected in the epithelium of the tubal mucosa, smooth muscle cells and the blood vessel endothelium. Western blotting showed that porcine oviducts contain 75, 48 and 45 kDa immunoreactive LH receptor proteins, like the corpus luteum (CL). The lowest receptor expression was found in controls and in gilts treated with estradiol or progesterone. Combined treatment with estradiol and progesterone resulted in a significant increase of LH receptor protein concentrations when compared with control animals. In vitro LH treatment affected oviduct contractility of combined estradiol and progesterone treated gilts but not the oviduct of the remaining groups. It also caused a decrease in amplitude, frequency and areas under the curve (AUC) of ampulla (P < 0.05) and the amplitude and AUC of isthmus (P < 0.001) in combined estradiol and progesterone-primed gilts. These results indicate that estradiol and progesterone together, but not separately, increase LH receptor protein in the porcine oviduct and that combined estradiol and progesterone priming is necessary for LH-induced relaxation of the porcine oviduct.     

Clinical evaluation of postpartum vaginal mucus reflects uterine bacterial infection and the immune response in cattle

Bacteria contaminate the uterus of most dairy cattle after parturition and endometritis causes infertility. An endometritis score can be ascribed based on the vaginal mucus character and odour but it is not clear if the clinical score reflects the number of uterine bacteria or the inflammatory response. The present study tested the hypothesis that clinical evaluation of endometritis reflects the number of bacteria present in the uterus, and the acute phase protein response. Swabs (n = 328) were collected from the uterine lumen of dairy cattle, 21 and 28 days postpartum, vaginal mucus was scored for character and odour, and blood samples collected for acute phase protein measurement. Bacteria were identified following aerobic and anaerobic culture, and the bacterial growth density was scored semi-quantitatively. When bacteria were categorised by their expected pathogenic potential in the uterus, purulent or fetid odour vaginal mucus was associated with the growth density of pathogenic bacteria but not opportunist contaminants. When bacteria were analysed independently, Arcanobacterium pyogenes, Proteus and Fusobacterium necrophorum growth densities were associated with mucopurulent or purulent vaginal mucus. The bacterial growth densities for A. pyogenes, Escherichia coli, non-hemolytic Streptococci, and Mannheimia haemolytica were associated with a fetid mucus odour. Peripheral plasma concentrations of alpha(1)-acid glycoprotein were higher if there was a fetid compared with a normal vaginal mucus odour (1.50 +/- 0.09 mg/mL versus 1.05 +/- 0.02 mg/mL, P < 0.001), but did not differ significantly between vaginal mucus character scores. The evaluation of the character and odour of vaginal mucus reflects the number of bacteria in the uterus, and the acute phase protein response.     

In vitro binding to and in vivo effect on the cytosol and nuclear progesterone receptors of various progestins,and their relationship to synthesis of uteroglobin in rabbit uterus

In vitro binding affinities of various progestins to cytosol and nuclear progesterone receptors of rabbit uterus were determined and correlated with the biological potency of these steroids. In addition, cytosol and nuclear progesterone receptor levels were measured after a 5-day administration of different progestins (0.5 mg/kg daily) with variable biologic activites. The receptor levels were compared with the bilological response; the induction of uteroglobin synthesis. Cytosol and nuclear progesterone receptors had identical steroid binding properties (r = 0.98). The correlation between the in vitro binding affinity (cytosol or nuclear) and the in vivo biologic activity of the steroids was good (r = 0.73). After a 5-day treatment with progestins, the nuclear receptor concentration correlated in an inverse manner (r = ?0.84) with the uterine fluid unteroglobin concentration. A similar, but slightly weaker correlation (r = ?0.81) was also found for the cytosol receptor content and uteroglobin secretion. These data indicate that not only nuclear, but also cytosol progesterone receptor levels decrease in the rabbit uterus during chronic hormone action. Decline in the nuclear progesterone receptor content seemed to occur during treatment with all progestational steroids, while onlyi progestins with high biological potency were capable of decreasing the cytosol receptor content.     

Dynamics of bacteriologic and cytologic changes in the uterus of postpartum dairy cows

The objectives of this study were to characterize clinical, intrauterine, bacteriologic and cytologic changes during the first month after parturition in healthy dairy cows and in cows with subclinical endometritis (SE) or clinical endometritis (CE). Furthermore, risk factors related to clinical bacteriologic and cytologic findings were determined. A total of 170 calvings were enrolled, and intrauterine samples were collected on Days 0, 3, 9, 15, 21, and 28 postpartum using the cytobrush technique. The presence of Escherichia coli and Trueperella pyogenes was determined by Fourier transform infrared spectroscopy. The cows were categorized according to their uterine health status (UHS) on Day 21 as healthy (clear or absent vaginal discharge and <5% polymorphonuclear cells [PMN] in the cytologic sample), SE (clear or absent vaginal discharge and ≥5% PMN), or CE (vaginal mucus containing any signs of pus). The prevalence of SE and CE on Day 21 was 27.9% and 58.4%, respectively. Generally, samples from cows with SE and CE showed a greater bacterial growth density (BGD) than those from healthy cows. The BGD tended to be affected by the interaction of time by UHS (P = 0.057). Differences between healthy, SE, and CE cows were found from Day 3 to the last sampling day. Furthermore, the percentage of PMN differed between healthy, SE, and CE cows and was affected by time in a cubic way (decrease/increase/decrease). Overall, E coli was found in 25.4% of the samples, and T pyogenes was identified in 30.2% of the samples. The risk for CE was increased by BGD and the presence of T pyogenes. Conversely, the presence of E coli had no effect on the risk of CE or the risk of SE. The risk for an infection with T pyogenes was greater in the first-parity cows and in cows with assisted calving. In conclusion, changes in BGD and proportion of PMN varied with the UHS (healthy, SE, and CE), which was affected by the presence of T pyogenes but not E coli.     

The involvement of luteinizing hormone (LH) and Pregnancy-Associated Glycoprotein family (PAG) in pregnancy maintenance in the pig

The paper presents the effect of in vivo immuno-neutralization of porcine luteinizing hormone (pLH) by species-homologous porcine antiserum (anti-pLH) administrations on pregnancy maintenance and immunodetection of the PAG proteins in precipitated plasma proteins of pregnant gilts. Pregnant gilts were passively immunized with 100 ml of porcine anti-pLH (titer 1:10 000) by multiple intravenous infusions performed from 37(th) to 42(nd) day post coitum (dpc; 12-h intervals). Blood samples of pregnant gilts were taken 12 times daily from 35 until 50 dpc. Concentrations of progesterone (P(4)) and pLH were determined by radioimmunoassays in systemic blood plasma of treated gilts and control pregnant gilts. The immuno-neutralization of peripheral pLH with the use of homologous anti-pLH serum resulted in a significant reduction (p<0.001) of plasma P(4) concentrations in two out of six treated gilts only, but abortion did not occur. In the remaining four passively immunized pregnant gilts, plasma P(4) concentration was increased (p<0.001) and the abortion occurred (47 dpc) only in one of the gilts. In addition, various anti-pPAG sera were purified by sequential adsorptions with endometrial proteins of cyclic gilts. Western blotting demonstrated the expression of the PAG proteins in precipitated plasma proteins of pregnant gilts. In conclusion, the passive immuno-neutralization of porcine LH by species-homologous antiserum (anti-pLH) did not affect the pregnancy maintenance. Thus, the maintenance of mid-pregnancy in gilts may depend also on other than LH luteotrophic factors. In addition, Western analysis of precipitated plasma proteins of pregnant pigs suggests a role of the PAG family during pregnancy in the pig.     

Pathologic Evaluation of Type 2 Porcine Reproductive and Respiratory Syndrome Virus Infection at the Maternal-Fetal Interface of Late Gestation Pregnant Gilts

The pathogenesis of fetal death caused by porcine reproductive and respiratory syndrome virus (PRRSV) remains unclear. The objective of this study was to improve our understanding of the pathogenesis by assessing potential relationships between specific histopathological lesions and PRRSV RNA concentration in the fetuses and the maternal-fetal interface. Pregnant gilts were inoculated with PRRSV (n = 114) or sham inoculated (n = 19) at 85±1 days of gestation. Dams and their litters were humanely euthanized and necropsied 21 days later. PRRSV RNA concentration was measured by qRT-PCR in the maternal-fetal interface and fetal thymus (n = 1391). Presence of fetal lesions was positively related to PRRSV RNA concentration in the maternal-fetal interface and fetal thymus (P<0.05 for both), but not to the distribution or severity of vasculitis, or the severity of endometrial inflammation. The presence of fetal and umbilical lesions was associated with greater odds of meconium staining (P<0.05 for both). The distribution and severity of vasculitis in endometrium were not significantly related to PRRSV RNA concentration in maternal-fetal interface or fetal thymus. Endometrial inflammation severity was positively related to distribution and severity of vasculitis in endometrium (P<0.001 for both). Conclusions from this study suggest that type 2 PRRSV infection in pregnant gilts induces significant histopathological lesions at maternal-fetal interface, but they are not associated with presence of PRRSV in the maternal-fetal interface at 21 days post infection. Conversely, fetal pathological lesions are associated with presence of PRRSV in the maternal-fetal interface and fetal thymus, and meconium staining is significantly associated with the presence of both fetal and umbilical lesions observed 21 days post infection.     

Nutrient restriction induces failure of reproductive function and molecular changes in hypothalamus–pituitary–gonadal axis in postpubertal gilts

People on a diet to lose weight may be at risk of reproductive failure. To investigate the effects of nutrient restriction on reproductive function and the underlying mechanism, changes of reproductive traits, hormone secretions and gene expressions in hypothalamus–pituitary–gonadal axis were examined in postpubertal gilts at anestrus induced by nutrient restriction. Gilts having experienced two estrus cycles were fed a normal (CON, 2.86 kg/d) or nutrient restricted (NR, 1 kg/d) food regimens to expect anestrus. NR gilts experienced another three estrus cycles, but did not express estrus symptoms at the anticipated fourth estrus. Blood samples were collected at 5 days’ interval for consecutive three times for measurement of hormone concentrations at the 23th day of the fourth estrus cycle. Individual progesterone concentrations of NR gilts from three consecutive blood samples were below 1.0 ng/mL versus 2.0 ng/mL in CON gilts, which was considered anestrus. NR gilts had impaired development of reproductive tract characterized by absence of large follicles (diameter ≥ 6 mm), decreased number of corepus lutea and atrophy of uterus and ovary tissues. Circulating concentrations of IGF-I, kisspeptin, estradiol, progesterone and leptin were significantly lower in NR gilts than that in CON gilts. Nutrient restriction down-regulated gene expressions of kiss-1, G-protein coupled protein 54, gonadotropin-releasing hormone, estrogen receptor α, progesterone receptor, leptin receptor, follicle-stimulating hormone and luteinizing hormone and insulin-like growth factor I in hypothalamus–pituitary–gonadal axis of gilts. Collectively, nutrient restriction resulted in impairment of reproductive function and changes of hormone secretions and gene expressions in hypothalamus–pituitary–gonadal axis, which shed light on the underlying mechanism by which nutrient restriction influenced reproductive function.     

Prostaglandin F2α back transfer from the mesometrium vasculature into the uterus of the gilt during early pregnancy and estrogen-induced pseudopregnancy

Isolated reproductive tracts from gilts on the days of luteal regression (13–17 day of estrous cycle), pregnant gilts (14–18 days of pregnancy) and estrogen-induced pseudopregnant gilts (15–18 day of estrous cycle) were supplied with autologous, oxygenated blood. ³H-PGF_2α (10⁸ dpm) was infused at a constant rate into three different sites of the most superficial layer of the myometrium along the length of the horn close to the broad ligament. During infusion (60 min) and 60 min after the infusion had been stopped, arterial blood was collected continuously in 5-min samples from a small branch of the uterine artery in the mesometrium area about 10 cm from the uterine horn. A significantly higher concentration of ³H-PGF_2α in the uterine-artery blood plasma was found in pseudopregnant and pregnant gilts than in the control group. The total ³H-PGF_2α back transfer with arterial blood from the broad ligament vasculature into the uterus was 2.4 × 10⁶ dpm, 6.0 × 10⁶ dpm and 19.8 × 10⁶ dpm of infused ³H-PGF_2α in the control, pregnant and pseudopregnant gilts, respectively. We suggest that ability for PGF_2α binding and back transfer from the broad ligament vasculature into the uterus, as observed in pseudopregnant and pregnant gilts, may strongly reduce the peak concentration during the pulsatile release of PGF_2α from the uterus and may protect the corpus luteum against luteolysis.     

Changes in prostaglandin production and ovarian function in gilts during endometritis induced by Escherichia coli infection

The aim of this study was to determine the prostaglandins (PGs) production and ovarian function in gilts after intrauterine infusions of 10(6) and 10(9) colony-forming units (cfu)/ml of Escherichia coli (E. coli). In Experiments 1 and 2, 30 ml of saline or 30 ml of E. coli suspension containing 10(6) or 10(9)cfu/ml, were infused once into each uterine horn in three groups of gilts on day 3 of the estrous cycle, respectively. In Experiment 1, 17 days after treatment it was revealed that inoculation of E. coli 10(9)cfu/ml induced severe acute or subacute endometritis while 10(6)cfu of E. coli evoked moderate acute endometritis or resulted in no inflammatory changes. In the gilts receiving 10(9)cfu/ml of E. coli, the concentration of 13,14-dihydro-15-keto-PGF(2)alpha in blood from the jugular vein was elevated (P<0.05-0.001) compared to concentration in the gilts inoculated with 10(6)cfu on days 8-17 after treatment. Both the E. coli-treated groups had a lower (P<0.05, P<0.01) progesterone plasma level from days 10 to 14 after administration than the control group. On day 17 of the study, infusion of E. coli 10(9)cfu/ml, in comparison to 10(6)cfu, resulted in the greater (P<0.001) content of PGE(2) in the myometrium. The content of both PGs in the endometrium as well as PGF(2alpha) in the myometrium of gilts-treated with 10(9)cfu/ml of E. coli was lower (P<0.001) than in gilts-treated with 10(6)cfu of bacteria. Newly formed corpora lutea were found in the gilts infused with 10(6), but not those infused with 10(9)cfu/ml of E. coli on day 17 after infusion. On day 8 of the study (Experiment 2), the blood from utero-ovarian vein of the gilts-treated with 10(9)cfu/ml of bacteria had a higher (P<0.05) PGF(2alpha) level and lower (P<0.001) PGE(2) level than following infusion of E. coli 10(6)cfu/ml. Also on day 8 of the study, the content of PGE(2) in the endometrium, both the PGs in the myometrium as well as cyclooxygenase-2 in the endometrium and myometrium was greater (P<0.01, P<0.001) after applying 10(9)cfu/ml than 10(6)cfu/ml of E. coli. These results indicate that intrauterine infusions of 10(6) or 10(9)cfu/ml of E. coli lead to the development of inflammatory states of different intensities which is connected with different PGF(2alpha) and PGE(2) production and function of ovaries.     

Bacterial inoculation speeds zinc release from ground tire rubber used as Zn fertilizer for corn and sunflower in a calcareous soil

Background and Aims

We tested the utility of some biological treatments to hasten degradation of waste tire rubber in soil and thus the release of zinc and sulfur for plant uptake.

Methods

Three rates of ground tire rubber (0, 150, and 300?mg?kg^?1) were incorporated into a Zn-deficient calcareous soil. Before addition to the soil, ground rubber was given four microbial treatments including no inoculation, inoculation with Rhodococcus erythropolis, inoculation with R. erythropolis+Escherichia coli, and inoculation with R. erythropolis+E. coli+Acinobacter calcoaceticus. In the pot experiment, corn (Zea mays L. Hybrid Single Cross 500) and sunflower (Helianthus annuus L. cv. Record) plants were exposed to three rates of ground rubber (0, 150, and 300?mg?kg^?1) or 3?mg zinc kg^?1 as ZnSO₄. Before addition to the soil, ground rubber and ZnSO₄ were inoculated or non-inoculated with R. erythropolis+E. coli+A. calcoaceticus.

Results

Ground rubber and microbial inoculation treatments reduced soil pH and the magnitude of this reduction increased over time. Ground rubber in combination with microbial inoculation increased DTPA-extractable soil Zn and Fe. The amount of DTPA-extractable Zn and Fe of rubber-amended soils increased over time so that the highest concentration of available Zn and Fe was found at week 10. Application of microbial inoculated ground tire rubber significantly increased shoot Zn concentration of each plant species.

Conclusions

Bacterial inoculation of ground rubber was effective in hastening increase in DTPA-extractable Zn in the studied calcareous soil and in enhancing Zn uptake by plants.     

Regulation of prostacyclin synthase expression and prostacyclin content in the pig endometrium

Prostaglandins (PGs) are critical regulators of a number of reproductive processes, including embryo development and implantation. In the present study, prostacyclin (PGI₂) synthase (PGIS) mRNA and protein expression, as well as 6-keto PGF_1α (a PGI₂ metabolite) concentration, were investigated in the pig uterus. Endometrial tissue and uterine luminal flushings were obtained on Days 4 to 18 of the estrous cycle and pregnancy. Additionally, conceptuses were collected and examined for PGIS mRNA expression and 6-keto PGF_1α concentration. Regulation of PGI₂ synthesis in the porcine endometrium by steroids, conceptus products, and cytokines was studied in vitro and/or in vivo. Endometrial PGIS protein level increased on Days 12 and 16 in pregnant but not in cyclic gilts. Moreover, higher PGIS protein expression on Day 12 of pregnancy was accompanied by a greater content of 6-keto PGF_1α in the endometrium. The concentration of 6-keto PGF_1α in uterine luminal flushings increased substantially on Days 16 and 18 in pregnant gilts and was higher than in cyclic animals. Greater PGIS mRNA expression and PGI₂ metabolite concentration were detected in Day 12 and 14 conceptuses, respectively. Incubation of endometrial explants with conceptus-conditioned medium resulted in upregulation of PGIS protein expression and increased PGI₂ secretion. Moreover, PGIS mRNA and protein expression were upregulated in the endometrium collected from gravid uterine horn on Day 14 of pregnancy. In summary, PGIS is differentially expressed in the endometrium of cyclic and pregnant gilts resulting in higher PGI₂ synthesis in pregnant animals. Porcine conceptuses are important regulators of endometrial PGIS expression and PGI₂ release during the implantation period.     

Relationship of length of uterus in prepubertal pigs and number of corpora lutea and fetuses at 30 days of gestation

Relationships of the length of the uterus at one reproductive stage to the length at other stages and the effect on potential litter size were determined. In Experiment 1, the length of the uterus was measured in situ at 20, 60, or 100 days of age at laparotomy with 20 gilts in each of the three age groups. Forty days after the initial measurement, the uterus was again measured in situ, gilts were ovariectomized and hysterectomized, and associations among uterine measurements at the two different stages were determined. Correlations between uterine length in situ and between uterine length and weight 40 days later were all greater than 0.75 (P < 0.001). The length of one uterine horn increased from 13.9 cm at 20 days of age to 36.7 cm at 140 days of age (P < 0.01). In Experiment 2, 66 gilts were unilaterally hysterectomized and ovariectomized (UHOX) at 150 days of age and the ovary was weighed. Length of the one horn was measured in 36 gilts. At 10 days after first estrus, the length of the remaining uterine horn was measured at laparotomy and corpora lutea were counted in 53 gilts. In 15 of the 53 gilts the remaining uterine horn was removed to obtain uterine weights. At the second or third estrus, 38 gilts were mated and at Day 30 of gestation, 31 gilts were pregnant. The gilts were killed, and the length of the uterus measured and corpora lutea (CL) and fetuses were counted. The length of one uterine horn at 150 days of age was 70 cm with a range of 47–110 cm. At 10 days after first estrus, length had increased to a mean of 141 cm with a range of 86–194 cm and at 30 days of gestation the mean was 244 cm with a range of 186–311 cm. There were 12.2 CL at first estrus, which was not different from 12.4 CL at the second or third estrus. The mean number of fetuses in one horn at 30 days of gestation was 9.5 with 77% prenatal survival. Length of uterine horn at 150 days of age was correlated with uterine horn length (r = 0.56, P < 0.001) at 10 days after first estrus and number of live fetuses (r=0.39, P<0.05) at 30 days of gestation. At Day 10 after first estrus, uterine length was not correlated with the number of CL, whereas at Day 30 of gestation, the number of CL and uterine length were correlated with the number of live fetuses in those gilts with below the mean number of live fetuses, but not in those gilts with above the mean of live fetuses. The number of live fetuses (r=0.66, P < 0.001) and fetal survival (r=0.63; P < 0.001) were correlated with uterine horn length in pregnant UHOX gilts. Length of the prepubertal uterus gives an indication of postpubertal length and the potential litter size in pigs.