The impact of zearalenone on the level of the selected estrogens in blood serum of sexually immature gilts

The aim of the study was to evaluate the influence of low dose (LOEL - lowest observed effect level) of zearalenone (200 microg/kg b.w.), applied per os for 7 days (short-term intoxication), on sexual behavior, concentration of the examined xenobiotic and its metabolite and selected estrogens in sexually immature gilts: ovariohysterectomised (group D1) and intact (group D2) animals. Clinical signs of oestrus (reddening, oedema and hyperaemia of the vulva and serorhoea from the reproductive tract--lack of standing reflex) were obserwed in group D1 on day 6 and in group D2 on day 4 of the experiment. Laboratory analyses of blood plasma were carried out determine the presence of zearalenone and alpha-zearalenole. They revealed an increase in the level of alpha-zearalenol before the oestrus, decrease in total amount of both examined substances on day when the oestrus appeared and increase in the level of both examined xenobiotics in the post oestrus period together with the higher share of zearalenone. Medium concentrations of estrone and estradiol within the borders of method determination in the majority of periods examined. Higher levels of estrone (32.0 pg/ml) were found on day 4, in the group D2 and estradiol (6.5 pg/ml) on day 6 in the D2 group. The presents study revealed that zearalenone applied per os at LOEL dose causes the incidence of apparent sexual readiness (without standing reflex) in sexually immature gilts with the somatically immature reproductive system.     

Morphology and ultrastructure of small intestine mucosa in gilts with zearalenone mycotoxicosis

Zearalenone (ZEA) is a mycotoxin produced by Fusarium. It evokes disruptions in sex cycle and changes in the morphology of the reproductive organs in pigs. The morphology and ultrastructure of the small intestine mucosa were assessed after the course of the experimental zearalenone mycotoxicosis. During the experiment, pigs were fed feed containing low doses of ZEA. The experiment lasted 7 days and it was carried out on sexually immature gilts that were divided into 3 groups: control group (C) and 2 experimental groups (E1, E2; n=8). Groups E1 and E2 were given ZEA every day at a dose of 200 and 400 microg/kg b.w. The samples of tissues were taken for examinations after the slaughter on the last day of the experiment. No significant changes in the morphology of the intestinal mucosa were found in the experimental animals. Increased activity of the goblet cells and high content of mucinogen vesicles, lymphocytic infiltration, increased activity of endocrine cells and the presence of Paneth cells on the bottom of intestinal crypts, which sugest mobilisation of the local defence mechanisms in the intestinal mucosa, were obseved in group E2.     

Bioactivation of zearalenone by porcine hepatic biotransformation

Zearalenone (ZEA) is a resorcylic acid lactone derivative produced by various Fusarium species that are widely found in food and feeds. The structure of zearalenone is flexible enough to allow a conformation able to bind to mammalian oestrogen receptors, where it acts as an agonist. Using oestrogen-dependent Human Breast Cancer (MCF-7) cells, the oestrogenic activity of zearalenone and its derivatives were compared using 17 beta-oestradiol as a positive control. The results obtained demonstrate that the oestrogenic potency of ZEA derivatives could be ranked in the following order: alpha-zearalenol > alpha-zearalanol > zearalenone > beta-zearalenol. Since pigs have been reported to be among the most sensitive animal species, biotransformation studies with pig liver subcellular fractions were conducted. These studies indicated that alpha-zearalenol is the main hepatic metabolite of zearalenone in pigs, and it is assumed that 3 alpha- and 3 beta-hydroxysteroid dehydrogeneases are involved in the hepatic biotransformation, since the formation of alpha-zearalenol and beta-zearalenol could be inhibited by prototypic substrates for either enzyme. The bioactivation of ZEA into the more active alpha-zearalenol seems to provide a possible explanation for the observed high sensitivity of pigs towards feeding-stuffs contaminated with the mycotoxin.     

Level of zearalenone in blood serum and lesions in ovarian follicles of sexually immature gilts in the course of zearalenone micotoxicosis

The aim of the study was to determine how a low dose of zearalenone applied orally for eight days influences the level of zearalenone (ZEN) and alpha-zearalenole in blood plasma and causes the occurrence of histopathological changes in the cells of the ovarian follicles in sexually immature gilts. The animals were divided into 2 groups (control, C; n = 4 and experimental, E; n = 4). The gilts from group E were treated daily with zearalenone at a dose of 200 microg/kg b.w. The level of zearalenone and alpha-zearalenole (ZON as the sum of the levels of both zearalenone and alpha-zearalenole) was measured daily. On day eight of the experiment the animals were sacrificed and their ovaries were taken for histopathological examination. The tissue sections obtained were HE- and PAS-stained according to McManus. The presence of PCNA antigen was also estimated. The highest concentration of ZON was noted on day 5 in group E (8.16 +/- 2.49 ng/ml). External estrus symptoms without standing reflex were observed in group E on day 4. In group C there were no pathological changes in the ovaries. In group E, a few ovarian follicles were found, but they were located in the cortical layer. They were filled with a liquid substance rich in protein and without the granulosa layer. There was disintegration with apoptotic-like changes of the PCNA-negative cells in the granulosa layer of single mature follicles. On day 4 the dose of zearalenone caused disturbances in the process of development and maturation of some of the best developed ovarian follicles. This probably occurred through the activation of on apoptosis-like process of the granulosa cells with simultaneous manifestation of estrus without standing reflex.     

玉米赤霉烯酮对断奶小母猪生产性能、血清抗氧化功能和免疫功能的影响

为了研究不同水平玉米赤霉烯酮(zearalenone,ZEA)污染饲粮对断奶小母猪生产性能、血清抗氧化功能、血清抗体水平及外周血淋巴细胞增殖率的影响。将40头健康三元杂交(杜×长×大)断奶小母猪按日龄[(35±1)日龄]和平均体重[(14.01±0.86)kg]分为4组,对照组饲喂基础饲粮(ZEA水平的测定值为0 mg/kg),试验组在基础饲粮中分别添加0.5、1.0及1.5 mg/kg ZEA[ZEA水平的测定值分别为(0.52±0.07)mg/kg、(1.04±0.03)mg/kg和(1.51±0.13)mg/kg]。预试期10 d,正试期35 d。结果表明:饲粮ZEA对断奶小母猪平均日采食量、平均日增重和料重比没有显著影响(P0.05)。与对照组相比,ZEA显著降低了血清谷胱甘肽过氧化物酶(GSH-Px,0.5、1.0和1.5 mg/kg ZEA)活性、猪瘟(1.0和1.5 mg/kg ZEA)和伪狂犬病病毒抗体水平(1.5 mg/kg ZEA)以及外周血淋巴细胞增殖率(1.0和1.5 mg/kg ZEA)(P0.05),而显著升高了血清丙二醛(MDA,0.5、1.0和1.5 mg/kg ZEA)含量(P0.05)。随着饲粮中ZEA水平的升高,断奶小母猪的料重比呈一次线性降低趋势(P=0.075),血清GSH-Px、超氧化物歧化酶(SOD)活性,血清病毒(猪瘟、伪狂犬病和高致病性猪蓝耳病病毒)抗体水平和外周血淋巴细胞增殖率均呈一次线性降低(P0.05),而血清MDA含量则呈一次线性升高(P0.05)。由此可见,饲粮中0.5 mg/kg的ZEA足以诱导小母猪的氧化应激反应,1.0 mg/kg的ZEA能够显著降低断奶小母猪的特异性体液免疫和细胞免疫功能。     

玉米赤霉烯酮中毒大鼠卵巢组织Bax和Bcl-2的表达

10周龄SD大鼠单剂量腹腔注射5mg／kg玉米赤霉烯酮玉米油溶液，分别于3、6、12、24、48h时剖杀取卵巢组织，检测不同时间卵巢组织Bax和Bcl-2的表达。结果显示，病理组织学观察发现卵巢组织出现不同程度损伤，卵泡颗粒细胞发生凋亡。免疫组化SP法试验组与对照组卵巢组织中均有Bax和Bcl-2的表达，并且随着时间的推进呈现动态变化。Bax在3、6、12h时表达量上调，表达量与对照组比较差异显著（P〈0．05），24、48h时表达量下降，与对照组比较差异不显著（P〉O．05）。3～48h卵巢组织中Bcl-2表达量与对照组比较差异不显著（P〉0．05）。结果表明，大鼠玉米赤霉烯酮中毒可引起卵巢组织的病变及颗粒细胞凋亡，且Bax在玉米赤霉烯酮中毒大鼠卵巢颗粒细胞凋亡中起着重要作用，Bcl-2的作用不明显。     

Metabolic profile of pigs fed feed containing zearalenone destructor

Zearalenone (ZEA) is a member of macrocyclic lactons family. It is a toxin--phytosteride produced by fungi of Fusarium ssp. genus. Zearalenone contaminates food and animal feeding stuffs and its destruction is difficult. It requires application of particular compounds that would bind zearalenone in the feed or feeding stuff or in the gastrointestinal tract and decrease its bio-accessibility. It should also fulfil all the safety requirements regarding the plant supplements and animals that are fed with this feed. The aim of the study was to estimate if the feed supplemented with different doses of zearalenone and zearalenone destructor causes changes of the metabolic profile in gilts. The results obtained show that applied destructor did not cause negative haematological and biochemical changes in the blood of the gilts examined. It can be suggested that it is a safe feed supplement pigs in prevention of zearalenone micotoxicosis.     

玉米赤霉烯酮的生殖毒性研究进展

玉米赤霉烯酮是一种特殊毒性的生物毒素,它的强雌激素样作用对动物机体的伤害很大,不仅引起动物流产、死胎、低产仔率等生殖机能异常,还可以导致繁殖功能下降、不育、畸形等.已有研究表明,ZEA对肝脏、肾脏、生殖系统和免疫系统产生明显的损伤,对细胞和遗传性也有毒性作用.目前,虽然国内外学者做了大量的研究,来揭示玉米赤霉烯酮损害机体生殖功能的机制,但关于它影响机体的生殖性能更详尽的机制还亟待进一步的研究.文章主要针对玉米赤霉烯酮的生殖毒性进行了综述,总结出ZEA对生殖系统的毒性机制,从而为临床预防及治疗繁殖疾病提供理论依据.     

Interaction of zearalenone and soybean isoflavone in diets on the growth performance, organ development and serum parameters in prepubertal gilts

The aim of the present research was to determine the interactive effect of zearalenone (ZEA) and soybean isoflavone (ISO) on the growth performance, development of organs and serum parameters in prepubertal gilts. Ninety 75-day-old female pigs (Duroc × Landrace × Yorkshire, 26.5 ± 0.60 kg) were randomly allocated to nine diet treatments during the 21-day study. The experiment employed a 3 × 3 factorial design using a non-soybean meal diet with the addition of 0, 0.5 or 2.0 mg/kg ZEA and 0, 300 or 600 mg/kg ISO. The results indicated that simultaneous addition of ZEA and ISO had no significant influence on the growth performance in prepubertal gilts. Zearalenone with 2 mg/kg increased (p < 0.05) the relative weight of the reproductive organs (including uterus and vagina) but had no obvious effects (p > 0.05) on the relative weight of the heart, liver, lung, kidney and spleen. Isoflavone at 600 mg/kg could offset the increased weight of the reproductive organs induced by ZEA. Simultaneous addition of ZEA and ISO to prepubertal gilts increased the level of alanine aminotransferase, aspartate aminotransferase and alkaline phosphatase in the serum (p < 0.05) at day 14 but their levels decreased (p < 0.05) over time. Zearalenone increased the level of malondialdehyde and decreased the concentrations of superoxide dismutase and glutathione peroxidase (p < 0.05) in the serum. The results suggested that ISO added to diets at 600 mg/kg could reduce the increase in the relative weight of reproductive organs and relieve the oxidative stress induced by ZEA added at 2 mg/kg during the growth phase in prepubertal gilts.     

玉米赤霉烯酮对断奶小母猪卵巢指数及孕激素受体分布和表达的影响

本试验旨在研究不同水平玉米赤霉烯酮(ZEA)污染饲粮对断奶小母猪卵巢指数及孕激素受体的阳性分布和mRNA相对表达量的影响。选择40头35~38日龄健康三元杂交(杜×长×大)断奶小母猪,按平均体重[(14.01±0.86)kg]随机分为4个组,采用单笼饲养。对照组饲喂基础饲粮,试验组分别在基础饲粮中添加0.5、1.0和1.5 mg/kg的ZEA,4个组ZEA的测定值分别为0、(0.52±0.07)mg/kg、(1.04±0.03)mg/kg和(1.51±0.13)mg/kg。试验期35 d。结果表明:随着饲粮ZEA水平的升高,断奶小母猪卵巢指数呈一次线性升高(P0.01);1.0 mg/kg ZEA组的卵巢指数显著高于1.5 mg/kg ZEA组(P0.05),1.5 mg/kg ZEA组的卵巢指数显著高于0.5 mg/kg ZEA组和对照组(P0.05)。随着饲粮ZEA水平的升高,断奶小母猪卵巢孕激素受体累积光密度(IOD)和mRNA相对表达量均呈一次线性升高(P0.01);1.5 mg/kg ZEA组的孕激素受体IOD和mRNA相对表达量显著高于1.0 mg/kg ZEA组(P0.05),1.0 mg/kg ZEA组又显著高于0.5 mg/kg ZEA组和对照组(P0.05)。免疫组化结果显示断奶小母猪卵巢中孕激素受体免疫阳性物质主要分布于原始卵泡和生长卵泡的卵母细胞及颗粒细胞、卵泡膜细胞及血管壁细胞。随着饲粮ZEA水平的升高和卵泡闭锁程度的增加,免疫阳性反应明显增强,但卵巢内孕激素受体免疫阳性物质的分布位置没有因ZEA的水平发生明显改变。由此可见,ZEA(1.0~1.5 mg/kg)可以通过调控断奶小母猪卵巢中孕激素受体的高水平表达,使孕激素受体的水平超出正向调节卵泡生长的限值,从而促使卵泡闭锁,降低卵巢指数,改变卵巢发育,进而影响断奶小母猪生殖系统的健康。     

A sensitive bioassay for detection of dietary estrogens in animal feeds

Estrogen-responsive proliferation in the MCF-7 cell line was used as a bioassay for detection of dietary estrogens. The bioassay procedure was adapted to screen for estrogenic activity in feedstuffs that have been associated with hyperestrogenism in livestock. Methanolic feed extracts were added to the cell culture medium at microliter/ml concentrations for 4 days, after which the cell proliferation response was measured as DNA content. The half-maximal response for estradiol occurred at 2 pM, or 0.54 pg/ml. For zearalenone, a weaker estrogen, the half-maximal response occurred at approximately 200 pM, or 64 pg/ml. The bioassay was calibrated against a number of known estrogens (estradiol, diethylstilbestrol, zearalenone, zearalanol [cattle implant], beta-zearalenol, zearalane), including the naturally occurring phytoestrogens (formononetin, genistein, daidzein, biochanin A, and coumestrol). The estrogenic activity of feed samples was expressed as equivalents of zearalenone (ppm zearalenone) that would have to be present to equally stimulate proliferation of the MCF-7 cells. The sensitivity of the bioassay was 0.05-0.1 ppm equivalents of zearalenone in feed, well below the threshold level associated with reproductive problems. The feed additive melengestrol acetate (MGA) showed no estrogenic activity in this assay. Estrogenic activity of feed extracts was confirmed by competitive inhibition with the antiestrogens tamoxifen or LY156758 (keoxifene) to show that stimulation of growth by feed extracts was through an estrogenic mechanism. Confirmation of known estrogens was by tandem mass spectroscopy. The assay is a sensitive and reliable screening procedure for detecting estrogenic activity in feedstuffs.     

咖啡酸对玉米赤霉烯酮诱导小鼠卵巢颗粒细胞凋亡的保护作用

卵巢颗粒细胞通过与卵母细胞相互作用及其自身分泌作用为卵泡的形成和发育成熟提供特殊的微环境。多种有害刺激可引起颗粒细胞凋亡和代谢失调从而降低卵母细胞质量,对胚胎的形成产生负面影响。玉米赤霉烯酮（zearalenone,ZEA）是畜禽养殖业中常见的造成卵巢颗粒细胞损伤的霉菌毒素,且缺少有效治疗药物。因此,本试验用ZEA造成小鼠卵巢颗粒细胞损伤,探究咖啡酸对ZEA诱导的小鼠卵巢颗粒细胞凋亡的保护作用。通过机械法分离小鼠卵巢颗粒细胞;运用间接免疫荧光法对分离出的细胞进行鉴定;使用MTT法测定咖啡酸对正常小鼠卵巢颗粒细胞活性的影响;选取200、100和50 μg·mL^-1咖啡酸分别与ZEA共处理颗粒细胞,同时设置细胞对照组和ZEA模型组,24 h后显微镜观察细胞形态及贴壁情况,MTT法检测细胞活力;qRT-PCR技术检测caspase-3 mRNA的表达;Western blot检测cleaved-caspase-3和cleaved-PARP蛋白水平。结果发现,FSHR阳性染色大量出现在试验组细胞胞浆中,提示分离到的细胞是小鼠卵巢颗粒细胞;咖啡酸对卵巢颗粒细胞没有毒性作用且细胞活力均在90%以上;与空白对照组细胞相比,ZEA组细胞体积较小,贴壁较差,细胞间隙增大,细胞活力显著降低（P<0.001）,caspase-3 mRNA相对表达量以及cleaved-caspase-3和cleaved-PARP蛋白表达水平显著升高（P<0.001）,而给予咖啡酸处理后细胞间隙减小,贴壁紧密,细胞活力显著升高（P<0.001）,显著降低ZEA诱导的caspase-3 mRNA含量增加（P<0.001）,显著降低凋亡相关蛋白cleaved-caspase-3和cleaved-PARP的表达（P<0.001）。本研究发现,咖啡酸可通过抑制ZEA诱导的细胞凋亡,恢复颗粒细胞活性。     

The Effect of Caffeic Acid on Zearalenone-induced Ovarian Granulosa Cell Apoptosis in Mice

Ovarian granulosa cells provide a special microenvironment for follicle formation and maturation through interaction with oocytes and their own secretion. A variety of harmful stimuli can cause granulosa cell apoptosis and metabolic disorders, reduce the quality of oocytes and have a negative impact on embryo formation. Zearalenone (ZEA) is a common cause of ovarian granulosa cells injury in the livestock industry, which is produced by mycotoxins, and lack of effective treatment drug. Therefore, in the current study zearalenone was used to induce ovarian granulosa cell injury and to explore the protective effect of caffeic acid on zearalenone-induced ovarian granulosa cell apoptosis in mice. Mouse ovarian granulosa cells were isolated by mechanical method, and indirect immunofluorescence was used to identify the isolated cells. MTT assay was used to determine the effect of caffeic acid on the activity of normal mouse ovarian granulosa cells.After granulosa cells were co-treated with caffeic acid (200, 100 and 50 μg·mL^-1) and ZEA for 24 hours, and control and ZEA group were set up at the same time, cell morphology and adherence were observed under a microscope. MTT was also used to detect cell viability. Caspase-3 mRNA expression level was detected by qRT-PCR. Cleaved-caspase-3 and cleaved-PARP protein expre-ssion levels were determined by Western blot. The results showed that positive FSHR staining appeared in cell cytoplasm of the test group, which confirmed that the isolated cells were mouse ovarian granulosa cells. The cell viability was above 90% which showed that caffeic acid had no toxic effect on granulosa cells. Compared with control group, ZEA group had smaller cell size, poor adherence, increased cell gap, and significant reduction in cell viability (P<0.001). Furthermore, the relative expression of caspase-3 mRNA, and cleaved-caspase-3 and cleaved-PARP protein level were significantly increased (P<0.001) compared with the control group. After caffeic acid treatment, cell gap was reduced, adherence was tight, cell viability was significantly increased (P<0.001). Caffeic acid significantly reduced zearalenone-induced increase in caspase-3 mRNA, and cleaved-caspase-3 and cleaved-PARP protein expression level (P<0.001). This study indicated that caffeic acid can restore granulosa cell viability by inhibiting ZEA-induced apoptosis.     

Preliminary evaluation of influence of zearalenone on cocultures of granulosa and internal theca cells of ovarian follicles in bitches in in vitro culture

Zearalenone (ZEA) is an undesirable substance in feed materials and feed of plant origin. It is an example of the micotoxin that causes disturbances in the functioning of the reproductive system. The wide range of plant compounds in pet food means that ZEA may frequently have a negative effect on pet reproduction. An assessment of the influence of ZEA on the granulosa and theca cells of the ovarian follicle in bitches in vitro was carried out. The co-culture of the ovarian follicles was incubated for 72 hours with the addition of 12.5 ng/ml and 25.0 ng/ml of ZEA. Numerous vacuoles in the cytoplasm of the granulosa cells were noted in the culture with the addition of 25.0 ng/ml of ZEA. Preliminary investigations suggest negative effect of ZEA on the granulosa cells in bitches in vitro.     

玉米赤霉烯酮的污染和残留及其作用机制

玉米赤霉烯酮(ZEA)是由镰孢真菌产生的类雌激素毒素.本文综述了ZEA的污染现状及其在动物组织中的残留现状,并且总结了ZEA的可能作用机制.     

玉米赤霉烯酮毒性机理研究进展

玉米赤霉烯酮是一种特殊毒性的生物毒素,具有类雌激素样作用,能引起动物流产、死胎、返情等生殖机能异常,还可以导致生长下降、免疫抑制、不育、畸形等.国内外学者做了大量的研究,来揭示玉米赤霉烯酮损害机体的机制.它主要通过影响机体的生殖性能,引起细胞凋亡、致畸、损伤DNA、氧化损害、影响免疫机能等机制,来影响动物与人类的健康.文章主要针对玉米赤霉烯酮的毒性机理进行综述.     

Characterization of effects of zearalenone in swine during early pregnancy

Mature gilts (n = 16) were hand mated and randomly assigned to 1 of 4 groups of 4 gilts each. Treated gilts had 108 mg of purified zearalenone added to their diet on postmating days (PMD) 2 to 6, 7 to 10, or 11 to 15. Control gilts were given the same diet without added zearalenone. On PMD 6, 10, and 15, control gilts had venous cannulas placed in the jugular vein, and blood samples were taken at 20-minute intervals for 4 hours before feeding and 4 hours after feeding. Samples were collected from treated gilts on the last day that zearalenone was consumed. Samples were analyzed for follicle stimulating hormone, luteinizing hormone (LH), and prolactin. Single blood samples were taken by venipuncture on PMD 8, 12, 16, 20, 24, and 28 and at euthanasia and were analyzed for serum concentration of progesterone and estradiol-17 beta. All gilts were euthanatized 30 to 32 days after mating, and fetal development was assessed. Three gilts that were given zearalenone on PMD 7 to 10 were not pregnant and had regressing corpora lutea on the ovaries at euthanasia. All other treated and control gilts were pregnant. Serum samples from treated gilts on PMD 10 and 15 had lower mean prolactin concentrations than did those from controls. The number of LH spikes were fewer (P less than 0.05) in gilts that were given zearalenone on PMD 15 compared with those in controls on PMD 15. Serum progesterone concentrations indicated that corpora lutea regressed between PMD 20 and 28 in nonpregnant gilts.(ABSTRACT TRUNCATED AT 250 WORDS)     

玉米赤霉烯酮的代谢、毒性及其预防措施

玉米赤霉烯酮(ZEA)是由镰孢属真菌产生的类雌激素毒素.由于气候条件不同,食品和饲料中ZEA浓度差异很大(从几个mg/kg至几千个mg/kg).本文综述了ZEA的吸收、代谢和生物转化机制,ZEA的生殖毒性、细胞毒性、免疫毒性和遗传毒性,并且从预防土壤中镰孢菌及其相关霉菌的污染、收获和收获后真菌毒素的控制、污染颗粒的物理...     

A morphological and immunohistochemical study of healthy and atretic follicles in the ovary of the sexually immature ostrich (Struthio camelus)

The morphology of healthy and atretic follicles in the ovary of the sexually immature ostrich was described in the present study. In addition, the distribution of the intermediate filaments desmin, vimentin and smooth muscle actin, in these ovarian follicles, was demonstrated. Healthy and atretic primordial, pre-vitellogenic and vitellogenic follicles were present in the ovaries of the sexually immature ostrich. Atresia occurred during all stages of follicular development. Atretic primordial and pre-vitellogenic follicles were characterized by the presence of a shrunken oocyte surrounded by a multilayered granulosa cell layer. Two forms of atresia (types 1 and 2) were identified in vitellogenic follicles. In the advanced stages of type 1 atresia the follicle was dominated by a hyalinized mass. In contrast, in type 2 atresia the granulosa and theca interna cells differentiated into interstitial gland cells. Positive immunostaining for desmin was observed in the granulosa cells of only healthy primordial and pre-vitellogenic follicles. Atretic primordial and pre-vitellogenic follicles were immunonegative for desmin. Vimentin immunoreactivity was demonstrated in the granulosa cells of all follicles except the vitellogenic atretic follicles. The results of the present study indicate that ovarian follicles in the sexually immature ostrich undergo a cycle of growth and regression, which is similar to that reported in other avian species. Furthermore, based on the results of the immunohistochemical study, it would appear that the distribution and immunostaining of intermediate filaments changes during follicular development and atresia.     

Evidence for the localization of porcine reproductive and respiratory syndrome virus (PRRSV) antigen and RNA in ovarian follicles in gilts

The pathogenesis of porcine reproductive and respiratory syndrome virus (PRRSV) infection in ovary was studied in sexually mature, cycling, nonsynchronized gilts infected with the PRRSV 16244B, a virulent field strain. Previous studies have shown that PRRSV can be isolated from ovaries and is transplacentally passed from gilts to the fetuses. The cause of infertility following PRRSV infection is not known. In this study, we identified the tropism of PRRSV in ovarian tissue from experimentally infected gilts in samples collected between 7 and 21 days postinfection (DPI). Tissues were collected and examined by virus isolation, in situ hybridization (ISH), immunohistochemistry (IHC), and double labeling to identify PRRSV-infected cell types. PRRSV was isolated in ovarian follicles at 7 days DPI. The IHC and ISH indicated that PRRSV-positive cells in ovaries were predominantly macrophages, which were numerous in atretic follicles. No evidence of infection and/or perpetuation of PRRSV in ova was observed, indicating that the female gonad is an unlikely site of persistence. No alteration of the normal ovarian architecture that would support a possible role of PRRSV infection in porcine female infertility was observed.