Cystic Degeneration in Porcine Ovaries - First Communication: Morphology of Cystic Ovaries, Interpretation of the Results

Contents: The morphological appearance of ovarian structures in 79 sows with ovarian cysts was examined. Sixty sows had been culled due to sterility; 19 sows had been repeatedly investigated clinically before slaughter. It was found that cystic ovaries without corpora lutea (CL) had the largest diameter and volume (151.3 cm³), in comparison to both cystic ovaries with CL (85.7 cm³) and normalovaries (16.9 cm³). The number of cysts on ovaries without corpora lutea was approximately twice as high as on the ovaries with CL (23.3 vs 12.1 cystslsow; p < 0.001). A combined form of degeneration consisting of large (> 1.5 cm in diameter) and small (1.5 em in diameter) cysts was found in 63% of the animals, whereas degeneration with only small cysts occured in 8% of the cases. Dark CL, interpreted as being functional, were mainly present in animals with up to 10 cysts per sow (24 out of 27 sows, p < 0.001); pale CL, interpreted as being weakly functional, were mainly found on ovaries with more than 10 cysts (19 out of 23 sows; p < 0.001). Based on macroscopical differentiation cyclic activity was found to be dependent to the number of cysts. Regular or irregular estrous cycles were observed in oligocystic animals (10 cysts/animal) with functional CL (58%). Up to 75% of the animals with pale or absent CL showed no estrous activity.     

奶牛卵巢囊肿的防治

卵巢囊肿分为卵泡囊肿和黄体囊肿两种。卵泡囊肿是由于卵泡上皮变性、卵泡壁结缔组织增生变厚、卵细胞死亡、卵泡液未吸收或增加而形成，其壁较厚。黄体囊肿是由于未排卵的卵泡壁上皮细胞黄体化而形成，或是正常排卵后由于某种原因黄体不足，在黄体内形成空腔，腔内聚积液体而形成。     

Effects of Twice-Weekly Follicular Punctures of Ovaries With or Without the Corpus Luteum on Follicular and Luteal Dynamics

We investigated the effects of twice-weekly follicular punctures of ovaries with or without corpus luteum (CL) on follicular and luteal dynamics. A cross-over design was used, with each cow (seven Japanese Black beef cows) being assigned to one of the three groups at 2-month intervals. Follicular punctures were performed twice weekly for three consecutive weeks until day 20 (oestrus = day 0). All visible follicles (diameter >3 mm) in the ovaries bearing CL (ipsilateral group) or those in the contralateral ovaries (contralateral group) were aspirated. As a control, all visible follicles in both ovaries were aspirated (bilateral group). Follicular development, CL formation and progesterone concentrations in each cow were monitored from days 0 to 30. Follicular growth profiles in the punctured ovaries during/after puncture treatment were similar, irrespective of the presence of follicles in the unpunctured ovary and the CL in the punctured or unpunctured ovaries. After puncture, two cows (28.6%) each in the ipsilateral and bilateral groups did not exhibit behavioural oestrus until day 30, whereas all cows in the contralateral group exhibited oestrus. CL growth and increase in progesterone concentrations after the last follicular puncture in the bilateral group were delayed when compared with those in the ipsilateral group. Our results indicate that the presence of follicles in the unpunctured ovary and the CL in the punctured or unpunctured ovaries does not significantly influence follicular growth in punctured ovaries during/after puncture treatment. However, follicular puncture in ovaries bearing CL may disturb or delay oestrus occurrence after treatment.     

乏情和发情初产母猪卵巢microRNA差异表达分析

为了探索卵巢microRNA （miRNA）在初产母猪生殖调控中的作用,本研究利用Illumina高通量测序技术检测了乏情和发情初产母猪卵巢miRNA的表达谱,并对表达量较高且差异显著的miRNA进行生物信息学分析。结果显示,本研究构建的两个small RNA文库共鉴定出503个miRNAs,其中已知的303个,新预测的200个。在已知的miRNA中,ssc-miR-10b在两个文库中的表达量最高,其次为ssc-miR-143-3p和ssc-miR-26a;在新预测的miRNA中,chr13_2637_mature在两个文库中的表达量最高,其次为chr8_9994_mature。与发情母猪相比,共有145个miRNAs发生显著变化（read counts>10,∣log₂（fold-change）∣>1）,上调114个,下调31个。在进一步筛选的31个表达量较高且差异显著的miRNAs（read counts>1 000,∣log₂（fold-change）∣>1）中,新预测的chr13_2585_mature上调倍数最高,且只在乏情母猪卵巢中表达。31个miRNAs共预测到7 388个靶基因,KEGG信号通路分析显示,有2 788个靶基因注释到了297个KEGG通路,前20个最富集的通路部分与生殖过程或生殖活动调控相关,表明这31个miRNAs参与了初产母猪的生殖调控。本研究结果丰富了猪miRNA数据资源,为进一步深入研究初产母猪的繁殖性能提供了理论依据。     

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.     

Follicular Development and Ovulation in Sows: Effect of hCG and GnRH Treatment

Follicular growth, chronology of ovulation and embryo morphology were compared in sows ovulating spontaneously and sows, in which the ovulation was attempted induced by hCG or GnRH.Indwelling catheters were placed on day 1 (weaning = day 0) in the ear veins of 18 sows, which were then randomly divided into 3 groups: a control group (N = 6), a group (N = 6) given 750 iu hCG (Physex®) im 76h after weaning (hCG group) and a group (N = 6) given 500 µg GnRH (Fertagyl®) im 76h (N = 3) or 100h after weaning (N = 3) (GnRH group). Follicular diameter and time of ovulation were monitored by ultrasonography every 4h from day 3 until ovulation or development of cysts by means of a sector scanner fitted with a 5.0/7.5 MHz multiangle probe. Heat detection was performed every 8h from day 3 until ovulation. On day 13, the sows were slaughtered, the number of corpora luteae (CL) was counted, and embryos were flushed from the uteri. The control group showed clear heat symptoms, and on day 3, the follicles were typically 3–7 mm and grew up to 7–10 mm over 2 days, where they remained for approximately 24h until ovulation took place 41h ± 9h after first sign of standing heat. The hCG group exhibited no signs of heat, and the follicles only reached 5–8 mm in diameter at time of ovulation, which occurred 40h ± lh after hCG-injection. The GnRH group exhibited inconsistent signs of heat, and the follicles reached a maximum size of 7–12 mm in diameter where they remained for more than 24h. Only 2 sows in this group ovulated within 84–92h after the GnRH injection, and development of bursa cysts and cystic follicles was a common finding. The average number of CL was 18.2 ±5.7 per sow (N = 16, range: 3–27) with no significant difference between the groups. Total embryo recovery was 79 ± 13 % with no significant difference between groups. The embryo diversity calculated as standard deviation of the maximum diameter was higher in the hCG group as compared with the control group.It is concluded that (1) transrectal ultrasonography can be used in sows for accurate assessment of follicular growth and ovulation; (2) the use of hCG results in lack of heat symptoms and reduced follicle size at the time of ovulation when injected 76h after weaning; (3) administration of a single injection of GnRH, if given before the first signs of heat, results in inconsistent heat symptoms and no or late ovulations.     

Expression of Vascular Endothelial Growth Factor Receptors in Bovine Cystic Follicles

Cystic follicles have excess fluid derived from blood flow in the theca interna of the follicle; therefore, the vasculature network is related to cystic follicle formation. Vascular endothelial growth factor (VEGF) is a potent stimulator of blood vessel permeability and angiogenesis. The aim of this study was to examine the expression of VEGF receptors proteins and mRNA in cystic follicles to elucidate the VEGF system in cystic follicles. The expression of protein for VEGF receptors; fms‐like‐tyrosine kinase‐1 (Flt‐1) and foetal liver kinase‐1 (Flk‐1) was detected by the immunohistochemical method. The mRNA expression of Flt‐1 and Flk‐1 in cystic follicles was determined by RT‐PCR. Concentration of oestradiol‐17β and progesterone in the follicular fluid of cystic follicles was determined using ELISA. Flt‐1‐ and Flk‐1 proteins were localized in granulosa and theca interna cells and endothelial cells of theca layers. The intensity of Flt‐1 and Flk‐1 immunoreaction was similar among cystic follicles with various ratios of oestradiol‐17β/progesterone concentrations. The expression of Flt‐1 and Flk‐1 mRNA was similar, regardless of the ratio of oestradiol‐17β to progesterone in follicular fluid. These results demonstrate that cystic follicles have both VEGF receptors in the granulosa and theca interna layers, which may be responsible for the increased permeability of microvessels, causing the accumulation of follicular fluid in cystic follicles.     

Management and Genetic Factors Affecting Fertility in Sows

Contents The purpose of this article is to summarize the current knowledge on genetic and management methods for improving fertility in sows. Fertilization rate, litter size and the interval between weaning and oestrus are traits that can be monitored on farms. These traits are heritable but can also be influenced by management. Age at puberty and the interval between weaning and oestrus are genetically linked ( r _g = 0.3) and a shorter weaning-to-oestrus interval is related to a longer duration of oestrus. Consequently, selection may be used to simplify detection of oestrus but because of variation in duration of oestrus between lines/breeds, farm-specific insemination strategies are needed. The direct boar effect on fertilization is small, probably due to the superabundance of sperm cells that is available in each dose of semen. Consequently, the use of a large number of sperm cells per dose is the best management tool for avoiding fertilization problems in sows. Health status of the boars, both for artificial insemination and natural mating, is of importance in order to produce a large number of sperm cells/doses of good quality per ejaculate. It is concluded that factors that influence fertility in sows, particularly prior to and during fertilization, are mainly related to oestrus and the optimal timing of insemination. Selection for adequate symptoms of oestrus, together with farm-specific insemination strategies are needed.     

Role of Cell Death Ligand and Receptor System on Regulation of Follicular Atresia in Pig Ovaries

Several hundred thousand primordial follicles are present in the mammalian ovary, however, only a limited number develop to the pre-ovulatory stage, and then finally ovulate. The others, more than 99%, will be eliminated through a degenerative process called 'atresia'. The endocrinological regulatory mechanisms involved in follicular development and atresia have been characterized to a large extent, but the precise temporal and molecular mechanisms involved in the regulation of these events have remained unknown. From many recent studies, it is suggested that the apoptosis in ovarian granulosa cells plays a crucial role in follicular atresia. Notably, death ligand–receptor interaction and subsequent intracellular signalling have been demonstrated to be the key mechanisms regulating granulosa cell apoptosis. In this review, we provide an overview of granulosa cell apoptosis regulated by death ligand–receptor signalling. The roles of death ligands and receptors [Fas ligand (FasL)–Fas, tumour necrosis factor (TNF)α–TNF receptor (TNFR), and TNFα-related apoptosis-inducing ligand (TRAIL)–TRAIL receptor (TRAILR)] and intracellular death-signal mediators [Fas-associated death domain protein (FADD), TNF receptor 1-associated death domain protein (TRADD), caspases, apoptotic protease-activating factor 1 (Apaf1), TNFR-associated factor 2 (TRAF2), and cellular FLICE-like inhibitory protein (cFLIP), etc.] in granulosa cells will be discussed.     

母猪饲粮中添加过氧化氢酶对母猪及仔猪生长性能和抗氧化能力的影响

本试验旨在研究母猪妊娠后期和哺乳期饲粮中添加过氧化氢酶(CAT)对母猪繁殖性能、抗氧化能力、饲粮养分消化率以及仔猪生长性能和抗氧化能力的影响。试验选取二元杂交(长白×大白)经产母猪160头,按母猪预产期、胎次体重、体况相近均衡分布的原则随机分为4组(对照组和试验A、B、C组),饲粮中CAT添加量分别为0、50、100、150U/kg,每组40头,每头母猪为1个重复,试验周期从妊娠第90天开始至仔猪断奶。结果表明:与对照组相比,饲粮中添加CAT对总产仔头数、活仔数、健仔数、初生窝重有提高但未达到显著水平(P>0.05);在母猪饲粮中添加CAT对降低哺乳仔猪腹泻率效果显著(P<0.05);在母猪妊娠第104天,与对照组相比,试验B和C组中血清总抗氧化能力(T AOC)分别提高60.03%和53.70%(P<0.05);试验B组中血清CAT活性较对照组、试验A和C组都有显著提高(P<0.05)。在母猪哺乳第14天,与对照组相比,试验B组中血清CAT活性显著提高(P<0.05);与对照组相比,试验B组初乳中T AOC提高119.10%(P<0.05);试验B和C组的初乳CAT活性较对照组分别提高86.24%和76.95%(P<0.05);各试验组在粗蛋白质消化率上与对照组相比差异均为显著(P<0.05);各试验组在粗脂肪消化率上与对照组相比差异均为显著(P<0.05)。综上,在母猪饲粮中添加CAT可以一定程度上提高母猪总产仔数、母猪妊娠第104天及哺乳第14天血清中T AOC、CAT活性、初乳中T AOC、CAT活性,母猪抗氧化能力在饲粮中添加100U/kgCAT时提升最明显;在母猪饲粮中添加CAT可以显著提高粗脂肪和粗蛋白质消化率。     

母猪不同生长阶段饲养管理技术

在实际生产中根据母猪的不同生产阶段,分为后备期、间情期、妊娠期、临产期、哺乳期和空怀期等几个阶段。为便于饲养管理和提高养殖效益,应按照各个时期的生理特点和对营养成分的需求采取不同的饲喂方式和配制全价的饲料,满足母猪对营养物质的需求,能保持适宜的体况和膘情,适时发情和配种,并生产出更多更优的仔猪,提高母猪繁殖力和养殖效益。该文分别叙述不同阶段母猪的饲养管理要点。     

影响母猪分娩率的因素分析

母猪生产的目的就是为了生产数量多、体重大、体质健壮的仔猪.在母猪数量一定的条件下,母猪的分娩率直接影响仔猪的生产数量,提高母猪的分娩率,已成为当今养猪场的关键管理环节.     

Lectin‐binding Sites on the Normal and Pathologic Uterus of Sows

The lectin‐binding pattern was compared in the normal and pathological uterus of sows during the ovarian cycle. The following biotinylated lectins were used: Con A, DBA, SBA, PNA, RCA‐I, UEA‐I and WGA. Glycoconjugate labelling showed differences between phases of ovarian cycle and presence of morphologic lesions. Cystic endometrial hyperplasia increased the RCA‐I reaction in the apical region of the glandular epithelium. There was higher intensity of labelling of WGA in the glandular epithelium in uteri with endometritis. In addition, increased Con A binding in the glandular epithelium and mild reduction of UEA‐I reactivity in the glycocalyx of the glandular epithelium were detected in the cases of endometritis. The results of this study show that morphologic alterations modify the sugar pattern in the porcine uterus. These modifications in glycoconjugates may be one of the reasons for decreased fertility in sows.     

Post-mortem Examination of Sows Genital Organs Culled for Reproductive Disturbances and Immunohistochemical Studies on ERα and PR A Receptors in the Anoestral Sows Uterus

The aim of present study was post-mortem examination of ovaries, uterus and plasma oestradiol-17beta (E2) and progesterone (P4) concentrations in the blood of sows with reproductive disturbances and the distribution of oestradiol receptor (ERalpha), as well as progesterone (PR-A) in the anoestrous sows uteri. Reproductive organs of 150 crossbred sows (Lithuanian White x Danish Landrace) culled for the reasons of reproductive disturbances, were collected in local abattoir over a period of 3 months (September-November). Organs were assessed to determine the stage of the oestrous cycle or anoestrus and their development. Blood samples were collected for E2 and P4 analysis from the jugular vein 1 h prior to slaughter. For this study uterine samples only from pathological anoestrous sows were subjected to immunohistochemical staining to assess the distribution of ERalpha and PR-A in surface epithelium, subepithelial connective tissue, glandular epithelium and myometrium. Macroscopic examination of the ovaries showed that 68.7% sows had active cycling ovaries, 26.6% sows were anoestrus, ovaries were small without CL, and 4.7% of sows had multiple follicular cysts. In anoestrous sows (n = 27) the number and intensity of the nuclear staining of ERalpha varied between different uterine tissue compartments. The highest number (>80%) and the strongest intensity (+++) of positively stained cells for ERalpha was seen in myometrium and glandular epithelium. In other uterine wall compartments the number and intensity of positively stained for ERalpha nuclei was lower (+/++). The PR-A was absent from all tissue compartments. The intensity of the nuclear staining for ERalpha varied not only between the different uterine compartments but also between the sows. The 11.1% of the sows presented ERalpha in surface epithelium, 74.1% of the sows in glandular epithelium and 63.0% of sows in the myometrium.     

Altered Expression of Transforming Growth Factor‐Beta Isoforms in Bovine Cystic Ovarian Disease

Cystic ovarian disease (COD) is one of the main causes of infertility in dairy cattle. It has been shown that intra‐ovarian factors may contribute to follicular persistence. Transforming growth factor‐beta (TGFB) isoforms are important paracrine and autocrine signalling molecules that regulate ovarian follicle growth and physiology. Considering the importance of these factors in the ovarian physiology, in this study, we examined the expression of TGFB isoforms (TGFB1, TGFB2 and TGFB3) in the ovary of healthy cows and animals with spontaneous and adrenocorticotrophic hormone (ACTH)‐induced COD. In the oestrous‐synchronized control group, the expression of TGFB1 in granulosa and theca cells was higher in spontaneous cysts than in atretic or tertiary follicles. When we compared TGFB2 expression in granulosa cells from atretic or tertiary follicles from the oestrous‐synchronized control group with that in ACTH‐induced or spontaneous follicular cysts, we found a higher expression in the latter. The expression of the TGFB isoforms studied was also altered during folliculogenesis in both the spontaneous and ACTH‐induced COD groups. As it has been previously shown that TGFB influences steroidogenesis, ovarian follicular proliferation and apoptosis, an alteration in its expression may contribute to the pathogenesis of this disease.     

Follicular Growth is Characterized by Biochemical Interplay of Proliferation and Apoptotic Death in Bovine Follicular Cells

Contents
This study was conducted to characterize the degree of proliferation and atresia in mid-luteal and follicular phase bovine follicles of different size containing morphologically good quality cumulus oocyte complexes. Follicular cell proliferation was estimated by enzyme immunoassay for tissue polypeptide specific antigen and atresia by an enzyme immunoassay for apoptosis. The relationship between follicular apoptosis and proliferation was inverse in the mid-luteal stage ovaries, whereas in follicular stage ovaries the tissue polypeptide specific antigen was increased in follicles with the highest degree of apoptosis. Follicular stage follicles showed a higher degree of apoptosis in comparison with the mid-luteal stage. This was caused by an increased apoptosis in follicles with less than 3 mm and greater than 5 mm diameter. In conclusion, the data indicate a sophisticated interplay between follicular growth and atresia in dependence of oestrous stage and follicular size.