哺乳期肉绵羊诱导发情及繁殖试验

为了使肉绵羊能在哺乳期发情怀孕,试验采用了在非繁殖季节,对24只处于哺乳期的试验羊用孕激素+孕马血清促性腺激素(PMSG)方法进行诱导发情处理.结果表明:其中12只羊发情,发情率为50.00%,发情时间为撤栓后2～3 d;对诱导发情成功的哺乳母羊进行配种,参加配种的12只羊有4只在预产期分娩产羔,其怀孕率仅为33.33%.     

哺乳期肉绵羊诱导发情及繁殖试验

为了使肉绵羊能在哺乳期发情怀孕,试验采用了在非繁殖季节,对24只处于哺乳期的试验羊用孕激素+孕马血清促性腺激素(PMSG)方法进行诱导发情处理。结果表明:其中12只羊发情,发情率为50.00%,发情时间为撤栓后2～3d;对诱导发情成功的哺乳母羊进行配种,参加配种的12只羊有4只在预产期分娩产羔,其怀孕率仅为33.33%。     

非繁殖季节母驴诱导发情、排卵效果的研究

[目的]研究前列腺素F2α（PGF2α）、孕马血清促性腺激素（PMSG）、人绒毛膜促性腺激素（hCG）3种激素在不同处理方案下对内蒙古通辽地区乏情期母驴卵泡生长情况、排卵率、发情率、受胎率与妊娠率的影响。[方法]将50头2～6岁、体重接近、体况良好、处于乏情期的母驴随机分为A、B、C、D4组,其中D组为未使用任何激素的空白对照组。首先,采用PGF2α、PMSG进行诱导发情试验,对A、B、C3组母驴注射2mL／头PGF2α,连续处理5d,第6天对A、B组母驴分别注射PMSG 1000IU,头和500IU,头;C组不做PMSG处理。诱导发情后将A、B组再细分为A1、A2和B1、B2组,当A1、B1、C3组中母驴卵泡直径大于35mm时注射hCG（总量1500IU）进行诱导排卵;A2、B2不做hCG处理,由卵泡自然发育至排卵。对所有诱导发情后有发情表现的母驴进行自然本交。同时,在试验期间利用直肠检查和B超检查对母驴的卵巢发育进行监测。『结果]c组发情率为30％,经PGF2α处理第5天时卵巢黄体基本全部消退;A组和B组的发情率分别为93．33％和86．66％,二者差异不大,但均明显高于C组。A1、B1最终的排卵率为100％和77．78％,而A2和B2的排卵率为60．00％和50．00％。A,组受胎率和妊娠率最高,分别达到了88．89％和80．00％,其次是B,组分别为66．67％和60．00％,B2、C2组的受胎率和妊娠率都较低,D组虽然有1头母驴发情,但配种后未妊娠。[结论]单独使用PGF2α对非繁殖季节母驴诱导发情效果不明显,但其对消除母驴黄体有明显作用;PGF2α与PMSG联用对诱导乏情母驴发情效果较好。使用PGF2α与PMSG对处于乏情期的母驴诱导发情后,注射hCG对母驴卵巢上优势卵泡具有明显的促进排卵作用,能加快优势卵泡的排卵,且明显提高母驴的排卵率。该试验确定的最佳方案为：注射0．2mgPGF2α连续处理5d,第6天注射PMSG 500lU,当优势卵泡直径达35mm时注射hCG 1500lU,排卵后配种。     

提高母猪繁殖性能的措施

１提高产仔数的措施１．１增加母猪排卵数实践证明，对空怀母猪在配种前进行短期优饲，有促进发情排卵和容易受胎的良好作用。另外，给母猪一次注射７５０～１５００IU的孕马血清（PMSG），并配合使用５００～１０００IU的人绒毛促性腺激素，可诱导母猪发情并刺激母猪超数排卵。１．２适时配种，提高排出卵的受精率由于品种、年龄及个体排卵时间的差异，因而在确定配种时间时，应灵活掌握。从品种来看，我国地方猪种发情持续时间较短，排卵较早，可在发情的第２d配种：引入品种发情持续期较长，排卵较晚，可在发情的第３～４d配种；杂种…     

应用激素诱导母羊发情的效果观察

东北细毛羊为短日照发情动物,1年1胎,除妊娠、哺乳时间外,1年中仍有4-5个月处于休情状态,每年的7-11月份才发情。为了使绵羊产羔后尽早发情、配种,提高生产效率,我们应用了CRID装置,PMSG促性腺激素和LRH-A3促性腺激素释放激素,复合孕酮制剂,对母羊进行诱导发情试验,并对发情配种效果进行了观察,取得了一些可靠数     

同期排卵-定时输精程序对后备母猪繁殖性能的影响

本研究旨在从配种次数、配种时间以及促排卵药物的使用3个方面对后备母猪同期排卵-定时输精技术方案进行优化。将后备母猪随机分成6组：(1)自然发情对照组：母猪自然发情、配种;(2)简式定时输精组：使用烯丙孕素性周期同步化后母猪自然发情配种;(3)传统定时输精组：母猪性周期同步化后,经过孕马血清促性腺激素和戈那瑞林处理使得卵泡发育和排卵同步化,无需查情,固定时间输精;(4)两点式查情定时输精组：在传统定时输精的基础上增加了注射促排卵药物时和配种后24 h 2个查情、配种时间点;(5)诱导发情促排实时输精组：性周期和卵泡发育同步化后,母猪发情时注射戈那瑞林促排卵,实时输精;(6)诱导发情实时输精组：母猪性周期和卵泡发育同步化处理后,实时输精。使用B超监测各组母猪卵泡发育和排卵情况,评价不同程序对母猪发情时间、排卵集中度、卵泡囊肿和繁殖性能的影响。结果显示：与简式定时输精组相比,注射外源促性腺激素改变了卵泡生长速度和排卵时间,缩短了最后一次饲喂烯丙孕素到发情的间隔时间,但是并没有改变发情-排卵间隔时间和排卵前卵泡大小。诱导发情促排实时输精组母猪妊娠率（93.52%vs 76.85%）和分娩率（8...     

应用激素诱导母羊发情的效果观察

东北细毛羊为短日照发情动物，1年1胎，除妊娠、哺乳时间外，1年中仍有4～5个月处于休情状态，每年的7～11月份才发情。为了使绵羊产羔后尽早发情、配种，提高生产效率，我们应用了CRID装置，PMSG促性腺激素和LRH-A3促性腺激素释放激素，复合孕酮制剂，对母羊进行诱导发情试验，并对发情配种效果进行了观察，取得了一些可靠数据，现将实验效果报告如下。     

绵羊繁殖季节同期发情与非繁殖季节诱导发情效果的研究

为了提高并稳定繁殖季节和非繁殖季节新疆地方绵羊的同期发情效果,本研究通过不同激素处理方案对发情率、发情集中时间等数据进行分析。结果表明:繁殖季节,海绵栓+PG法、海绵栓+PMSG法与三者联合使用均可获得较好的发情效果(92.62%～96.96%,P0.05);发情集中于撤栓后的36～48 h,此阶段的发情率占总发情率的74.19%。而在非繁殖季节,使用海绵栓+PG的同期发情处理方法,发情率仅为56.12%,极显著低于海绵栓+PG+PMSG处理组的发情率90.43%(P0.01);发情也集中于撤栓后的36～48 h,此阶段发情率占总发情率的82.8%;非繁殖季节,每只绵羊注射PMSG 333 IU的发情率显著低于注射500 IU组和666 IU组(P0.05),且666 IU组的发情率最高。     

诱导青年母猪卵泡发育及排卵效果

应用孕马血清促性腺激素(PMSG)制剂对43头青年母猪进行诱导排卵,研究了其卵泡发育及排卵规律,并探讨了诱导处于卵巢机能静止期的母猪发情的最佳途径.结果表明,母猪接受交配时间越长,处理后的排卵效果越好,发情母猪初次交配时间集中在注药后69～126.5 h;卵泡发育总数为(11.90±4.41)个/头,与发情微弱及不发情母猪存在极显著差异(P<0.01),说明卵泡发育总数与母猪发情表现之间存在密切关系;进一步观察青年母猪早期胚胎发育的渐进性,发现进入输卵管内的胚胎处于2～4细胞期,进入子宫时处于4～8细胞期,时间在初配后51～59 h;胚胎全部进入子宫的时间在母猪初配后71～83.5 h,且诱导排卵不影响胚胎的质量.     

海门山羊超数排卵,卵巢反应和胚胎发育观察

用促性腺激素 FSH(280IU 和340IU)、PMSG(1000IU)对海门山羊进行超数排卵处理,并比较它们的超排效果.试验结果表明:用280IU 的 FSH 超排,排卵数平均为7.5±4.6;用340IU 的 FSH 超排,排卵数平均为8.6±6.2;用 PMSG 超排,排卵数平均为5.5.FSH 处理超排效果比 PMSG 处理效果好。FSH 组的两种剂量间的效果差异不显著(P>0.05);卵巢对激素的反应比较明显,卵泡多,卵巢体积增大.经观察,胚胎发育在发情配种后的第3天,以单细胞受精卵到2细胞阶段的胚胎趋多;第4天,以4—8细胞阶段的胚胎趋多;第5天,以16—32细胞阶段趋多;第6天,发育到桑椹胚;第7—8天,以囊胚趋多。这一结果,为海门山羊今后进行胚胎工程提供了参考价值。     

Feline embryo transfer during the non-breeding season

To obtain normal kits by embryo treansfer (ET) during the non-breeding season, maintenance of pregnancy was carried out by administration of sustained action progesterone (P4) in queens. Embryos were recovered six days after mating from five donor queens in which ovulation was induced by administration of eCG and hCG. The number of embryos recovered ranged from 24 to 53 (mean: 37.2 +/- 6.4) per animal and most embryos were compacted morulae. The yield of embryos was 49.0-93.3% (mean: 73.8 +/- 9.6%). As for recipients, porcine pituitary gland preparation and hCG were administered to 19 queens and estrus and ovulation were induced in 18 queens (94.7%). These queens underwent intrauterine ET of five compacted morulae and 17 cats (94.4%) were impregnated. The number of implantations was 2-5 (mean: 3.7 +/- 0.3). Among these impregnated queens, 15 cats received P4 adminstration starting on day 24 of gestation and 1-5 newborns (mean: 3.4 +/- 0.3) were obtained by normal delivery or caesarean section on day 64-69 of gestation. However, two animals that were not treated with P4 underwent spontaneous abortion about the mid gestational period. Therefore, it is possible to obtain normal kits from queens in the non-breeding season by ET with maintenance of pregnancy by P4 administration.     

Synchrony of Ovulation and Follicular Dynamics in Merino Ewes Treated with GnRH in the Breeding and Non-breeding Seasons

The effect of gonadotropin releasing hormone (GnRH) treatment on the time of ovulation and the occurrence of follicular dominance during the non-breeding and breeding seasons (experiment 1), and on fertility after artificial insemination (AI) in the non-breeding season (experiment 2), was examined in Merino ewes. Oestrus was synchronized in 40 nulliparous ewes (experiment 1; n = 20, in the non-breeding and breeding seasons) and in 79 multiparous ewes (experiment 2) using intravaginal sponges and pregnant mare serum gonadotropin. Thirty six hours after sponge removal (SR), half the ewes were injected (i.m.) with 40 microg of synthetic GnRH and the remainder used as controls. GnRH improved the synchrony of ovulation compared with the controls in the breeding (SD = 2.8 vs 5.7 days, p = 0.04) but not the non-breeding season (SD = 3.8 vs 4.4 days, p = 0.69), with ewes ovulating from 42 to 54 h (mean 50.4 +/- 4.08 h) and 42-60 h (mean 54.4 +/- 5.47 h) after SR for GnRH and control, respectively. For both treated and control ewes, ovulation occurred earlier in the non-breeding than the breeding season (50.1 vs 54.6 h; p = 0.002). GnRH had no effect on follicular dominance, as assessed by divergence (D: the time the ovulatory follicle exceeded the average size of the other non-ovulating follicles) or on the interval from D to ovulation (IDO). However, follicular dynamics differed between seasons. The mean follicle diameter increased at a faster rate up to 36 h after SR in the non-breeding compared with the breeding season and then rapidly declined, compared with a later peak (42 h after SR) in mean follicular size during the breeding season. IDO was shorter in the non-breeding than in the breeding season (26.7 +/- 4.30 h vs 39.6 +/- 4.53 h; p = 0.05). In experiment 2, ewes (n = 38 GnRH-treated, n = 40 controls) were inseminated in the uterus by laparoscopy 42 h or 48 h after SR with frozen-thawed sperm. The fertility of ewes treated with GnRH (nine of 39, 23%) was not different to the controls (eight of 38, 21%; p = 0.01). In conclusion the application of GnRH improved synchronization of ovulation but did not improve fertility rates after AI.     

人绒毛膜促性腺激素对母驴促排卵效果的研究

[目的]研究人绒毛膜促性腺激素（human chorionic gonadotropin,hCG）对母驴卵泡发育、排卵率、受胎率以及血清生殖激素水平的影响。[方法]选择优势卵泡直径在30~35 mm以及大于35 mm的母驴各30头,不同优势卵泡直径的母驴群体分别设置1个500 IU/头hCG处理组（n=10）、1个1 000 IU/头hCG处理组（n=10）、1个不接受hCG处理的对照组（n=10）。采用肌肉注射方法对各组母驴进行hCG处理。每隔24 h进行1次B超检查,观察各组母驴卵泡发育情况,测量卵泡直径;记录各组发生排卵的母驴数量,计算排卵率。对各组母驴进行人工输精,输精后第18天进行孕检,记录各组受胎母驴头数,计算各组受胎率。于hCG处理后0、24、48、72 h分别测定各组母驴血清中雌二醇（estradiol,E₂）和孕酮（progesterone,PROG）水平。[结果]2个群体母驴的卵泡直径随hCG注射剂量的增加而增大;优势卵泡直径大于35 mm的母驴群体中,肌肉注射hCG的2个组在处理后24 h内均出现排卵,而对照组母驴没有排卵;优势卵泡直径不同的2个母驴群体,在hCG处理48 h后排卵母驴数和排卵率与对照组相比均有所提高,其中,hCG处理后72 h,优势卵泡直径大于35 mm的母驴群体中,1 000 IU/头 hCG处理组的排卵率达到100%。2个母驴群体中,接受hCG处理的母驴,受胎率均高于对照,并且随hCG剂量的增加,受胎率有所提高;优势卵泡直径大于35 mm的母驴群体中,1 000 IU/头 hCG处理组的受胎率达到50%。2个母驴群体中,1 000 IU/头处理组在hCG处理后24 h的血清E₂浓度均较0 h时有较大幅度的提升,在0~72 h内血清PROG浓度的总体提升幅度较大。[结论]hCG处理可提高母驴的排卵率、受胎率以及血清中E₂和PROG水平,1 000 IU/头剂量的效果更好。     

Observations on the influence of exogenous gonadotrophins and cloprostenol on ovulation in gilts

We studied the effects of gonadotrophins and prostaglandin (PG) F_2α on ovulation in gilts. Twenty-eight gilts were induced to ovulate using 750 IU pregnant mares serum gonadotrophin (PMSG) and 500 IU human chorionic gonadotrophin (hCG), administered 72 h apart. At 34 and 36 h after hCG, gilts received injections of either 500 μg or 175 μg PGF_2α (cloprostenol), or had no injections. Laparotomies were performed at 36 h (cloprostenol gilts) or 38 h (controls) after hCG injection. The ovaries were examined and the proportion of preovulatory follicles that had ovulated (ovulation percent) was determined at 30 min intervals for up to 6 h. The number of gilts in which ovulation was initiated and the ovulation percent increased (p<0.001) with time, but was not affected by treatment. Many medium sized follicles (≤6 mm) were also observed to ovulate, or to exhibit progressive luteinization without overt ovulation, during the surgical period. A discrepancy between numbers of preovulatory follicles and corpora lutea suggests that luteal counts may not be an accurate assessment of ovulation rate following gonadotrophic stimulation.     

Effect of hCG on Early Luteal Serum Progesterone Concentrations in PG600‐Treated Gilts

Gilt oestrus and ovulation responses to injection of a combination of equine chorionic gonadotrophin (eCG) and human chorionic gonadotrophin (hCG) (PG600) can be unpredictable, possibly reflecting inadequate circulating LH activity. The objective of this study was to determine the effect of PG600 followed by supplemental hCG on gilt ovarian responses. In experiment 1, 212 Hypor gilts (160 day of age) housed on two farms in Spain received intramuscular (i.m.) injections of PG600 (n = 47), or PG600 with an additional 200 IU hCG injected either concurrently (hCG‐0; n = 39), or at 24 h (hCG‐24; n = 41) or 48 h (hCG‐48; n = 45) after PG600. A further 40 gilts served as non‐injected controls. Ovulation responses were determined on the basis of initial blood progesterone concentrations being <1 ng/ml and achieving >5 ng / ml 10 d after the PG600 injection. The incidence of ovulating gilts having progesterone concentrations >30 ng/ml were recorded. During the study period, 10% of control gilts ovulated whereas 85–100% of hormone‐treated gilts ovulated. There were no significant differences among hormone groups for proportions of gilts ovulating. The proportions of gilts having circulating progesterone concentrations >30 ng/ml were increased (p ≤ 0.02) in all hCG treated groups compared with the PG600 group. In experiment 2, a total of 76 Hypor gilts at either 150 or 200 days of age were injected with PG600 (n = 18), 400 IU eCG followed by 200 IU hCG 24 h later (n = 20), PG600 followed by 100 IU hCG 24 h later (n = 17), or 400 IU eCG followed by 300 IU hCG 24 h later (n = 21). Blood samples were obtained 10 days later for progesterone assay. There were no effects of treatment or age on incidence of ovulation, but fewer 150‐day‐old gilts treated with PG600 or 400 IU eCG followed by 200 IU hCG had progesterone concentrations >30 ng / ml. We conclude that hCG treatment subsequent to PG600 treatment will generate a higher circulating progesterone concentration, although the effect is not evident in older, presumably peripubertal, gilts. The mechanism involved and implications for fertility remain to be determined.     

Human chorionic gonadotropin (hCG): the effect of dose on ovulation and pregnancy rate in Thoroughbred mares experiencing their first ovulation of the breeding season

AIM: To determine the effect of hCG dose on ovulation and pregnancy rate in Thoroughbred mares experiencing their first ovulation of the breeding season. METHODS: Over 3 successive breeding seasons, a total of 101 mares were randomly assigned to 1 of 4 treatment groups (intravenous injection of either saline, 1500, 3000, or 6000 IU hCG), as they approached their first ovulation of the breeding season. Mares were bred 1 day post-injection to 1 of 11 stallions, and every other day until ovulation occurred. Data were analysed using multivariable logistic regression with correction for over-dispersion due to clustering. RESULTS: Mares treated with hCG were more likely to ovulate within 72 h of treatment than mares treated with saline (p<0.001); there was no significant difference between doses of hCG on risk of ovulation (p>0.15). Farm also had a significant impact on the risk of ovulation (p=0.027). Mares treated with hCG were more likely to be diagnosed pregnant 14 days post ovulation than saline-treated mares (p=0.081, p=0.029 and p=0.026 for the 1500, 3000 and 6000 IU doses, respectively); there was no significant difference between doses of hCG on risk of pregnancy (p>0.45). CONCLUSIONS: A single injection of hCG (1500-6000 IU) is effective at inducing ovulation in late transitional mares and increases the likelihood of pregnancy at 14 days post ovulation. This paper supports the use of hCG as an integral part of optimal broodmare management.     

Effect of P.G. 600 on the timing of ovulation in gilts treated with altrenogest

We previously reported that ovulation rate, but not pregnancy rate or litter size at d 30 after mating, was enhanced by treatment with P.G. 600 (400 IU of PMSG and 200 IU of hCG, Intervet America, Inc., Millsboro, DE) in gilts fed the orally active progestin, altrenogest (Matrix, Intervet America, Inc.) to synchronize estrus. We hypothesized that in addition to increasing ovulation rate, P.G. 600 may have altered the timing of ovulation. Therefore, mating gilts 12 and 24 h after first detection of estrus, as is common in the swine industry, may not have been the optimal breeding regimen, and as a consequence, pregnancy rate and litter size were not altered. The objective of the present study was to determine the effect of P.G. 600 on the timing of ovulation in gilts treated with altrenogest. Randomly cycling, crossbred gilts (5.5 mo old, 117 kg BW, and 14.7 mm of backfat) were fed a diet containing altrenogest (15 mg/d) for 18 d. Twenty-four hours after altrenogest withdrawal, gilts received i.m. injections of P.G. 600 (n = 25) or saline (n = 25). Gilts were checked for estrus at 8-h intervals. After first detection of estrus, transrectal ultrasonography was performed at 8-h intervals to determine the time of ovulation. Gilts were killed 9 to 11 d after the onset of estrus to determine ovulation rate. All gilts displayed estrus by 7 d after treatment with P.G. 600 or saline. Compared with saline, P.G. 600 increased (P = 0.07) ovulation rate (14.8 vs. 17.5, respectively; SE = 1.1). The intervals from injection to estrus (110.9 vs. 98.4; SE = 2.7 h; P < 0.01) and injection to ovulation (141.9 vs. 128.6; SE = 3.2 h; P < 0.01) were greater in gilts treated with saline than in gilts treated with P.G. 600. Duration of estrus (54.4 vs. 53.7; SE = 2.5 h), the estrus-to-ovulation interval (30.2 vs. 31.7; SE = 2.2 h), and the time of ovulation as a percentage of estrus duration (55.8 vs. 57.5; SE = 3.0%) did not differ for the P.G. 600 and saline-injected gilts, respectively. In summary, P.G. 600 advanced the onset of estrus and ovulation following termination of altrenogest treatment and increased ovulation rate; however, treatment of gilts with P.G. 600 had no effect on the timing of ovulation relative to the onset of estrus.     

The effect of time of insemination with fresh cooled transported semen and natural mating relative to ovulation on pregnancy and embryo loss rates in the mare

One hundred and fifty-four mares were inseminated with fresh semen either during the pre- or post-ovulatory periods at different intervals relative to ovulation: 36-24 h (n = 17) and 24-0 h (n = 30) before ovulation; 0-8 h (n = 21), 8-16 h (n = 24), 16-24 h (n = 48) and 24-32 h (n = 14) h after ovulation. All mares received the same routine post-mating treatment consisting of an intrauterine infusion with 1 litre of saline and antibiotics followed 8 h later by an intravenous administration of oxytocin. Artificial inseminations (AI) from 36 h before ovulation up to 16 h post-ovulation were performed with transported cooled semen. While there was no data available for inseminations later than 16 h, data from natural mating after 16 h post-ovulation were included. Pregnancy rate (PR) of mares inseminated 36-24 h (29.4%) was significantly lower (p < 0.05) than mares inseminated 24-0 h before ovulation (60%), 0-8 h (66.7%) and 8-16 h (70.1%) post-ovulation. Embryo loss rate (ELR) was highest in mares mated 24-32 h after ovulation (75%). PR of mares mated 16-24 h post-ovulation (54.1%) did not differ significantly from any other group (p > 0.05); however, the ELR did increased markedly (34.6%) compared with inseminations before 16 h post-ovulation (<12%). At ≥ 30 days post-ovulation, PR of mares mated 16-24 h after ovulation (35.4%) was significantly lower than mares mated 0-16 h after ovulation (62%). Good PR with acceptable ELR can result from inseminations within 16 h of ovulation, at least with this specific post-mating routine treatment.     

Comparison of the effect of cronolone sponges and PMSG or cloprostenol on estrous induction in Turkish Saanen goats

The efficiency of cronolone sponges in combination with either pregnant mare serum gonadotrophin (PMSG) or cloprostenol (PGF2alpha) for inducing and synchronizing the estrous cycle in Turkish Saanen does was investigated during the transition from non-breeding to breeding season. All does (n = 80) were treated with 20 mg cronolone sponges for 11 days and divided into 4 equal groups. In addition, each doe received an intramuscular injection of either 1.5 ml sterile saline solution, 0.075 mg PGF2alpha, 500 IU PMSG or 500 IU PMSG and 0.075 mg PGF2alpha, 24 h before the sponge removal. Cervical artificial insemination (AI) with frozen-thawed semen was performed once 16 h after the detection of the first accepted mount. The total estrous response for the first 24 +/- 4 h, total estrous response within 96 h, time to onset of the induced estrus, duration of the induced estrus and pregnancy rate was found to be 75.0%, 97.5%, 31.4 +/- 1.2 h, 29.3 +/- 1.2 h, and 33.3%, respectively. There were significant differences between the first two groups and the last two groups in terms of the onset of induced estrus and estrous response at the first 24 +/- 4 h (P < 0.05). These results indicate that the use of cronolone/PMSG was more effective than cronolone/PGF2alpha in the attainment of early and compact induction of estrus in Turkish Saanen does.