Influence of zearalenone on reproductive system cell proliferation in gilts

Zearalenone (ZEA) is a macrocyclic lactone, estrogenic, diet-depending and fusaric micotoxin, which is produced on many kinds of cereals and feeds in the favourable conditions of humidity and temperature. The structure of ZEA is similar to the structure of estrogens and it enables binding to the estrogenic receptors. The stimulation of protein synthesis in the cells of the reproductive system, which causes intensification of cell proliferation, is one of the effects of ZEA actions. Oedema and vulva reddening are the clinical, external signs of ZEA intoxication in pigs. The aim of this study was to designate the degree of reproductive cell proliferation after low doses of ZEA were applied per os in sexually immature gilts with simultaneous monitoring of zearalenone and alpha-zearalenol levels in peripheral blood. The following were observed in the gilts examined fluctuations of zearalenone and alpha-zearalenol levels in blood, which were connected with entero-hepatic circulation and also numerous histopathological changes in ovarian follicle structure. These changes were present in the reproductive system of sexually immature gilts with a big contribution of PCNA-positive cells. The studies show that zearalenone application in sexually immature gilts caused ovarian follicle atresia and apoptoso-like changes in granule cells. Intensified cell proliferation, which was expressed with the growth of PCNA index, was observed in uterus and oviduct.     

Effects of zearalenone (F2) on estrous activity and reproduction in gilts

The effects of zearalenone on swine reproduction were investigated in two trials involving a total of 82 gilts which were allotted into three groups at puberty, mated at second estrus and slaughtered 80 d postbreeding. A control diet without mycotoxin (group 1) or an experimental diet containing 3.61 ppm (first trial) or 4.33 ppm zearalenone (second trial) were fed at a mean daily level of 2 kg/animal. The experimental diet was fed from puberty to mating (group 2) or during pregnancy (group 3). No difference was observed between the two trials. When fed to nonpregnant gilts, zearalenone induced a pseudopregnancy state in 45% of the animals; no estrus was detected within 50 d following puberty and corpora lutea developed at puberty were maintained. The uterine horns were edematous. Reproductive performance measured at 80 d postmating (ovulation rate, weight of corpora lutea, number of normal and abnormal fetuses, embryonic mortality) were not affected by zearalenone intake. But when zearalenone was fed during pregnancy, weights of uterus, placental membranes and fetuses were significantly decreased in comparison with those of control gilts and heterogeneity of fetuses in the same litter was increased. Hematocrit and erythrocyte count were lower in fetuses from gilts ingesting zearalenone, but hematology of the dams remained unaffected. No mycotoxin residue could be detected in gilts or fetal tissues despite the great consequences observed on cyclicity of the females or on the development of embryos. This experiment showed evidence of the estrogenic properties of zearalenone in mature gilts.     

Consequences of different patterns of feed intake during the estrous cycle in gilts on subsequent fertility

The impact of different patterns of feed restriction between d 1 and 15 of the estrous cycle on subsequent reproductive performance of 23 trios of littermate gilts was tested. Some gilts were fed a high plane of nutrition (HH gilts) throughout the cycle, in contrast to HR gilts, which were restricted from d 8 to 15, and RH gilts, which were restricted from d 1 to 7. During feed restriction, weight gain in RH gilts (2.5 +/- .7 kg) was lower (P = .006) between d 1 and d 7 than in their HH and HR littermates (5.6 +/- .7 and 5.6 +/- .8 kg, respectively) and it was lower (P = .0001) in HR gilts (5.5 +/- .5 kg) between d 8 to d 15 than in their HH and RH counterparts (8.5 +/- .4 and 9.4 +/- .5 kg, respectively). There were no differences in backfat changes among groups. Embryonic survival in HR gilts at d 28 of gestation (68.3 +/- 4.8%) was lower (P < .05) than in HH and RH gilts (83.6 +/- 4.3 and 81.7 +/- 4.5%, respectively). Plasma progesterone concentrations in HR gilts were lower (P < .05) at 48 and 72 h after onset of standing estrus (.82 +/- .2 and 3.6 +/- .5 ng/mL, respectively) than in HH and RH gilts (1.44 +/- .2 and 1.24 +/- .2 ng/mL, 5.0 +/- .4 and 5.0 +/- .5 ng/mL, respectively at 48 and 72 h). No differences in ovulation rate were observed among treatments. Placental area was positively correlated to embryo size at d 28 (embryo size = .0003 x (area) + 18.35; r = .28, P = .03) but placental volume was negatively correlated to the number of embryos in utero (placental volume = -4.317 x (number) + 207.55, r = -.39, P = .002). These data demonstrate that the timing of feed restriction during follicular development has important consequences for subsequent embryo survival, possibly mediated by differences in progesterone concentrations in early pregnancy.     

Comparative study of stress response,growth and development of uteri in post‐weaning gilts challenged with zearalenone and estradiol benzoate

The objective of this study was to evaluate the effects of zearalenone (ZEA) and estradiol benzoate (EB) on stress injury and uterine development in post‐weaning gilts. Thirty healthy post‐weaning female gilts (Duroc × Landrace × Large White) aged 28–32 days were randomly allocated to three treatments as follows: (a) basal diet (Control), (b) basal diet plus 1.0 mg/kg purified ZEA (ZEA) and (c) basal diet plus 0.75 ml (1.5 mg) EB per pig at 3‐days intervals by intramuscular injection (EB). The serum estradiol (E₂), the final and the increased vulvar area, uterine index, thickness of the myometrium and endometrium, and protein expression of heat shock protein 70 (HSP70) in ZEA group were higher than those in the control group (p < .05), but lower than those in the EB group (p < .05). The serum luteinizing hormone in ZEA group was lower than that of the control group (p < .05), but higher than that in the EB group (p < .05). Higher serum follicle‐stimulating hormone and progesterone were observed in the ZEA and control groups than those in the EB group (p < .05). The serum glutathione peroxidase activity in the ZEA group was lower than that in the control and EB groups (p < .001), and the malondialdehyde in the ZEA group was higher than that in the control and EB groups (p < .001). Moreover, the relative mRNA and protein expression of growth hormone receptor (GHR) and relative mRNA expression of HSP70 in the ZEA and EB groups were higher than those in the control group (p < .05). In conclusion, both ZEA (1.0 mg/kg) and EB (1.5 mg at 3 days intervals by intramuscular injection) stimulated vulvar swelling and uterine hypertrophy by disordering serum hormones and up‐regulating GHR expression, and induced stress by different mechanisms in this study. Furthermore, the observed up‐regulating HSP70 expression challenged by ZEA or EB may be part of the mechanism to resist stress injury.     

Results of histochemical and biochemical studies of uterus of gilts during the estrous cycle]

Alkaline phosphatase activity as well as lipid, polysaccharide, glycogen, and acid mucopolysaccharide levels in the uteri of 79 gilts were histochemically tested during the first three sexual cycles. Intra-cyclic alkaline phosphatase activity exhibited clearly pronounced variations in the surface epithelium, with maximum values reached in metoestrus, and moderate variations in the endometrial stroma, with maximum values reached in dioestrus. Cycle-dependent variations were recordable also from the glycogen and lipid levels in the surface and glandular epithelia and from the acid mucopolysaccharide levels in the endometrial stroma. The activity of alkaline phosphatase as well as the levels of glycogen and acid mucopolysaccharides during the first cycle still were lower than those in the second and third. Biochemical examinations of the endometrium revealed significant cycle-dependent variations in the activities of alkaline phosphatase, inorganic pyrophosphatase, and succinate-dehydrogenase as well as in the concentrations of soluble protein and glycogen, with maximum values being recordable on the tenth day of cycle. No significant intracyclic variations were recordable from the activities of acid phosphatase as well as of aspartate alanine-amino transferase.     

Production of estradiol by each ovary during the estrous cycle of cows

The objective of our experiment was to examine changes in serum concentrations of estradiol in each utero-ovarian vein before, during and after gonadotropin surges. Four cows were given prostaglandin F2 alpha (PGF2 alpha) during diestrus and three cows were allowed to cycle spontaneously. All cows had a cannula in each utero-ovarian vein and in one jugular vein. Most cows had two transient rises in estradiol, primarily coming from a single ovary, preceding and after luteinizing hormone (LH) surges. The first rise in estradiol began after luteal regression and was sustained from 48 h before a pre-ovulatory LH surge to the end of the LH surge. The second rise in estradiol was sustained from 72 to 168 h after the end of an LH surge. To determine how rapidly asymmetrical production of estradiol began during luteolysis, several cows were injected with PGF2 alpha during the luteal phase. Blood samples were taken from a jugular and both utero-ovarian veins at hourly intervals before and after PGF2 alpha. Asymmetrical production of estradiol began within 3 h after an injection of PGF2 alpha. We concluded: (1) that a single ovary was responsible for the sustained increases in concentration of estradiol that occur during proestrus to estrus and early diestrus in cows and (2) that cows may have at least one follicle capable of producing estradiol during most days of an estrous cycle, thus little delay in selection of which follicle eventually ovulates occurs after luteal regression.     

The effects of recombinant porcine somatotropin on reproductive function in gilts treated during the finishing phase.

The objective of this study was to determine the effects of recombinant porcine somatotropin (rpST) treatment during the finishing phase on subsequent reproductive function in crossbred gilts. Forty gilts weighing 50 kg and housed in a swine finishing facility were randomly assigned to control or rpST treatment. Four control and four rpST-treated gilts were allotted per pen. Twenty rpST-treated gilts received 6 mg of rpST.gilt-1.d-1 in 1 ml of buffered carrier and 20 control gilts received 1 ml of buffered carrier.gilt-1.d-1. Injections were administered daily at 1400 in the extensor muscle of the neck. All gilts received an 18% CP diet containing 1.2% lysine. Treatment was terminated when the average weight in each pen reached 110 kg. Gilts treated with rpST gained more weight (P less than .05) than control gilts (59.8 +/- 1.0 vs 53.5 +/- 1.0 kg). Age at puberty was not different (rpST, 182.2 +/- 3.3; control 181.4 +/- 3.1 d). Prior treatment with rpST did not significantly affect length of estrus (rpST, 1.9 +/- .1; control, 1.8 +/- .1 d) or estrous cycle length (rpST, 20.6 +/- .4; control, 20.4 +/- .4 d). Ovulation rates at second estrus were similar for rpST gilts (15.1 +/- .5) and control gilts (14.4 +/- .5). More embryos (P = .10) were recovered on d 9 to 12 of gestation from rpST-treated gilts than from control gilts (13.1 +/- .9 vs 10.7 +/- .9).(ABSTRACT TRUNCATED AT 250 WORDS)     

Prepubertal exposure to dietary zearalenone alters hypothalamo-hypophysial function but does not impair postpubertal reproductive function of gilts

At an average age of 70 d, 60 Yorkshire gilts born either in July (Trial 1; n = 30) or August (Trial 2; n = 30) received a diet containing zearalenone for 0 (control), 45 or 90 d. The concentration of zearalenone in diets was 2 ppm for 2 wk and 1.5 ppm for the remainder of the study. Vulval swelling and reddening was evident within 7 d after zearalenone was first fed. Zearalenone consumption had no effect on BW or backfat depth. Puberty occurred in Trial 1 at 219 +/- 6 d and was not influenced by zearalenone. Gilts in Trial 2 were divided into two groups; blood samples were taken from 12 gilts to assess pulsatile LH patterns and LH response to estradiol benzoate (EB) and 18 were handled similarly to those in Trial 1. Of this latter subgroup, age at puberty was younger (P less than .05) with zearalenone (217 +/- 7.0, 193 +/- 9.1 and 185.6 +/- 8.2 d for 0-, 45-, and 90-d treatments). Prepubertal consumption of zearalenone did not affect conception rates, ovulation rates, number of fetuses or percentage of embryo survival following mating at pubertal estrus. Two days before the 90-d experimental period ended for Trial 2, blood samples were taken from 12 gilts (four/treatment) every 15 min for 4 h prior to injection of EB (10 micrograms/kg) and every 6 h for 108 h after EB. Analysis of pulsatile patterns of LH revealed no influence of zearalenone on the number of peaks/4 h, baseline concentration or peak height.(ABSTRACT TRUNCATED AT 250 WORDS)     

Plasma gonadotropins and ovarian hormones during the estrous cycle in high compared to low ovulation rate gilts

Mature gilts classified by low (12 to 16 corpora lutea [CL], n = 6) or high (17 to 26 CL, n = 5) ovulation rate (OR) were compared for plasma follicle-stimulating hormone (FSH), luteinizing hormone (LH), progesterone, estradiol-17beta, and inhibin during an estrous cycle. Gilts were checked for estrus at 8-h intervals beginning on d 18. Blood samples were collected at 8-h intervals beginning on d 18 of the third estrous cycle and continued for one complete estrous cycle. Analysis for FSH and LH was performed on samples collected at 8-h intervals and for ovarian hormones on samples collected at 24-h intervals. The data were standardized to the peak of LH at fourth (d 0) and fifth estrus for the follicular phase and analyzed in discrete periods during the periovulatory (-1, 0, +1 d relative to LH peak), early-luteal (d 1 to 5), mid-luteal (d 6 to 10), late-luteal (11 to 15), periluteolytic (-1, 0, +1 d relative to progesterone decline), and follicular (5 d prior to fifth estrus) phases of the estrous cycle. The number of CL during the sampling estrous cycle was greater (P < 0.005) for the high vs low OR gilts (18.8 vs 14.3) and again (P < 0.001) in the cycle subsequent to hormone measurement (20.9 vs 14.7). For high-OR gilts, FSH was greater during the ovulatory period (P = 0.002), the mid- (P < 0.05) and late-luteal phases (P = 0.01), and tended to be elevated during the early-luteal (P = 0.06), but not the luteolytic or follicular periods. LH was greater in high-OR gilts during the ovulatory period (P < 0.005), but not at other periods during the cycle. In high-OR gilts, progesterone was greater in the mid, late, and ovulatory phases (P < 0.005), but not in the follicular, ovulatory, and early-luteal phases. Concentrations of estradiol-17beta were not different between OR groups during the cycle. Inhibin was greater for the high OR group (P < 0.005) during the early, mid, late, luteolytic, and follicular phases (P < 0.001). The duration of the follicular phase (from last baseline estrogen value to the LH peak) was 6.5 +/- 0.5 d and was not affected by OR group. These results indicate that elevated concentrations of both FSH and LH are associated with increased ovulation rate during the ovulatory phase, but that only elevated FSH during much of the luteal phase is associated with increased ovulation rate. Of the ovarian hormones, both inhibin and progesterone are highly related to greater ovulation rates. These findings could aid in understanding how ovulation rate is controlled in pigs.     

富铜酵母对西门塔尔牛发情周期生殖激素分泌的影响

选用16头18月龄、体况良好、体质量390 kg的西门塔尔牛育成母牛,采用完全随机分组设计分为4组,研究富铜酵母(0、8、16、24 mg/kg)对发情周期外周血清促黄体素、促卵泡素、孕酮和雌二醇分泌规律的影响.结果显示,8 mg/kg组和16 mg/kg组显著提高了发情周期促黄体素、促卵泡素、孕酮和雌二醇水平(P<0.05);24 mg/kg组有提高趋势,但与对照组比差异不显著(P>0.05).结果表明,添加富铜酵母以8～16 mg/kg对发情周期生殖激素分泌有促进作用,以8 mg/kg为佳,兼顾基础日粮的含铜量,建议日粮铜水平为13.87 mg/kg.     

Effect of purified zearalenone on early gestation in gilts

Purified zearalenone (Z) was added to the diet of gilts from d 2 to 15 postmating. Gilts received either 0, 5, 15, 30, 60 or 90 ppm Z (three to five gilts per dose) in 1.8 kg of feed daily. Serum concentrations of progesterone and estradiol-17 beta were determined weekly. On d 13 to 15 and 40 to 43 postmating, blood samples were drawn from a cannula at 20 min intervals for 4 h and analyzed for luteinizing hormone (LH), follicle-stimulating hormone (FSH) and prolactin (PRL). Gilts were killed 40 to 43 d postmating and embryonic development was assessed. Treatment with 5, 15 or 30 ppm Z had no effect on embryonic development when compared with 0 ppm. No fetuses were present in gilts fed 60 to 90 ppm Z, but two gilts given 60 ppm Z had remnants of fetal membranes in the uterus. The histologic appearance of reproductive tract tissues from the gilts given 60 ppm Z was similar to that from pregnant gilts. Tissues from gilts given 90 ppm Z appeared to be stimulated by both estrogen and progesterone. Serum concentrations of progesterone were decreased at 2, 3 and 6 wk postbreeding in gilts fed 60 and 90 ppm Z. Serum concentrations of estradiol-estradiol-17 beta were decreased at 4 wk postbreeding in gilts fed 60 and 90 ppm Z.(ABSTRACT TRUNCATED AT 250 WORDS)     

Induction of short-term pseudopregnancy in gilts by the administration of estradiol benzoate or estradiol dipropionate to achieve ovulatory synchronization for embryo collection

A study was conducted to investigate whether ovulation in gilts could be synchronized for embryo collection by the administration of estradiol benzoate (EB) or estradiol dipropionate (EDP) to induce pseudopregnancy, followed by the treatment with prostaglandin F_2α (PGF_2α) on 10 days after. Ten gilts each received a total of 20 mg of EB or EDP on Day 10 or EB on Day 10 and 14 to induce pseudopregnancy (Day 0 = onset of estrus). Donors received PGF_2α 10 or 15 days (as a control) after the first administration of estrogens and subsequently eCG and hCG, and were then inseminated artificially. The embryos were collected 7 days after the administration of hCG, and assessed for embryo yield and their developmental stages. All protocols resulted in good embryo yield (9.8–13.2 embryos in average), and the embryos showed average ability to develop to the expanded blastocyst stage (3.29–4.03 as developmental scores) without any significant differences among the protocols. These results suggest that the administration of PGF_2α 10 days after the treatment of gilts with EB or EDP would allow synchronization of ovulation and embryo collection, as well as shortening the period from estrus detection to embryo collection, thus improving embryo collection efficiency.     

Effect of levamisole on immune function and reproductive performance in first-litter gilts

First-litter commercial cross-bred gilts were treated with levamisole (1.5, 2.5, or 3.5 mg/kg of body weight) weekly during the last 4 weeks of gestation, because similar treatment of dairy heifers had improved postpartum maternal health and neonatal survival. In the gilts, differences in reproductive performance were not found on the basis of pig survival at birth, pig survival at weaning, birth weight, or weaning weight. Also, differences between treated and control gilts were not found in response of circulating lymphocytes to mitogen stimulation (phytohemagglutinin A, concanavalin A, and pokeweed mitogen). In all gilts, the lymphocyte response to mitogen stimulation was decreased during the first week after farrowing.     

玉米赤霉烯酮对雄性小鼠生殖系统的毒性作用

为探讨玉米赤霉烯酮(ZEA)对雄性生殖系统的毒性作用,本试验选取清洁级昆明系雄性小鼠93只,随机分成5组:25 mg/kg、50 mg/kg、75 mg/kg ZEA处理组、溶剂对照组(DMSO)、空白对照组(生理盐水),进行腹腔注射染毒,每天1次,连续5 d,于末次染毒后第2天摘眼球采血并脱颈处死动物,分离睾丸和附睾,测定脏器系数、精子密度和活力,睾丸酶的活力水平以及血清睾酮和雌激素的浓度。结果显示,与对照组相比,玉米赤霉烯酮显著地降低小鼠的体重和精液品质,睾丸乳酸脱氢酶(LDH)、碱性磷酸酶(AKP)、酸性磷酸酶(ACP)的活力随着染毒剂量的增加显著升高(P0.01,P0.05),血清睾酮和雌激素浓度随染毒剂量的增加显著下降(P0.01,P0.05)。该结果提示,玉米赤霉烯酮对雄性小鼠生殖系统具有严重的损伤作用,影响生精功能。     

The influence of male pheromones on the contractile reactivity of the isolated superficial veins of the nose and face during the estrous cycle in gilts

This study was designed to determine whether steroid sex pheromones of the boar may affect the contractile activity of the superficial venous vessels of the nose and face in gilts, and in this way contribute to recently discovered humoral transfer of pheromones to the brain and hypophysis. The dependence between the reactivity of nasal and facial veins to male pheromones and the phase of the estrous cycle in gilts was also studied. The gilts were used in the luteal phase of the cycle and in the follicular phase of the cycle. The dorsal nasal, frontal and facial veins were isolated on an appropriate day of the estrous cycle. The isolated rings of veins were treated with androstenone (5alpha-androst-16-en-3-one), androstenol (5alpha-androst-16-en-3-ol) and testosterone (17beta-hydroxy-4-androsten-3-one) in concentrations of 1 or 10 microM. Changes in the contractile activity of the isolated vein segments were measured using isometric transducer and recorded on HSE-ACAD software for Windows. Androstenone--main signaling boar pheromone--induced much stronger reactions of the vessels than androstenol. Androstenone caused significant relaxation of the dorsal nasal and frontal veins, and significant increased tension of the facial vein in the follicular phase of the estrous cycle. The results obtained suggest a direct effect of male pheromones on the contractile reactivity of the superficial veins of the nose and face in the female, and in this way contribute to a humoral pathway for transfer of pheromones to the brain and hypophysis. Moreover, the present study revealed changes in the reactivity during the estrous cycle of the veins, transporting blood from the region of the nasal cavity, to male pheromones participating in the regulation of female reproductive processes.