Epigallocatechin-3-gallate supplementation can improve antioxidant status in stressed quail

1. Heat stress causes oxidative stress, which decreases plasma antioxidants in poultry. Epigallocatechin gallate (EGCG), the most abundant polyphenol in green tea, is a powerful antioxidant against lipid peroxidation. This experiment was conducted to evaluate the effects of dietary EGCG supplementation on performance, carcase characteristics, concentrations of malondialdehyde, lipid peroxidation indicator, vitamins C, E, A, cholesterol, triglyceride, and glucose in Japanese quail (Coturnix coturnix japonica) exposed to a high ambient temperature of 34 degrees C. 2. One hundred and eighty Japanese quails (10 d old) were assigned at random to 6 treatment groups consisting of 10 replicates of three birds. Birds were kept in cages in a temperature-controlled room at either 22 degrees C (thermo-neutral) or 34 degrees C (heat stress) for 8 h/d. Birds were fed either a basal diet or the diet supplemented with 200 or 400 mg of EGCG/kg of diet. 3. EGCG supplementation linearly increased feed intake, live weight gain, feed efficiency, cold carcase weight and yield under heat stress conditions but did not show the same effect at thermoneutral conditions. Serum vitamin C, E, and A concentrations increased in birds reared at high temperature while non-significant changes occurred in thermo-neutral groups. Malondialdehyde concentrations in serum and liver decreased in all birds of both thermo-neutral and heat stressed groups as dietary EGCG supplementation increased. Heat stress-induced increase in serum cholesterol, triglycerides and glucose concentrations were linearly reversed by EGCG supplementation. 4. The results indicate that EGCG supplementation improved the live performance and antioxidant status of heat-stressed Japanese quail.     

Gossypol,a polyphenolic aldehyde from cotton plant,interferes with swine granulosa cell function

Gossypol is a polyphenol isolated from the seed, roots and stem of cotton plant (Gossypium sp.) It has been associated with adverse effects on female reproduction, but recently also shown having promising effects against several malignancies. Its mechanisms of action are however still not fully understood. This study was therefore conducted to investigate the effect of 5 or 25 μg/mL gossypol on swine granulosa cell steroidogenic activity, redox status and Vascular Endothelial Growth Factor (VEGF) production. Study demonstrated that gossypol significantly (P < 0.001) inhibited granulosa cell estradiol 17β and progesterone production, an effect that could be at least partially mediated by an increase (P < 0.05) of nitric oxide and superoxide anion production as a consequence of superoxide dismutase inhibition. Moreover, gossypol stimulates (P < 0.001) VEGF production. In conclusion, study has demonstrated effecs of gossypol on swine granulosa cell function in vitro. Effects on female swine fertility can not be excluded.     

The impact of the phyto‐oestrogen genistein on swine granulosa cell function

Soya and soybean products used in swine feeding contain genistein, a non‐steroidal phyto‐oestrogen which has been demonstrated to influence endocrine functions. This observation leads us to design this study to evaluate the effect of genistein on swine granulosa cell steroidogenesis and proliferation. In the attempt to unravel the genistein signal transduction mechanisms, we verified the effect of lavendustin, a Tyrosine Kinase (TK) inhibitor, and the potential involvement of NO/cGMP pathway. Finally, as angiogenesis is essential for follicle development, we tested the effect of the phyto‐oestrogen on vascular endothelial growth factor production and on granulosa cell redox status, because free‐radical species modulate neovascularization. Our data provide evidence that genistein interferes with granulosa cell steroidogenesis while it does not modulate cell growth: this effect could be at least partially produced by inhibiting TK‐dependent signalling systems. On the contrary, NO/cGMP pathway or vascular endothelial growth factor production can be excluded as signalling mechanism involved in phyto‐oestrogen effects. Remarkably, genistein stimulates hydrogen peroxide production thus potentially inhibiting follicular angiogenesis. Collectively, these results suggest that genistein consumption could potentially negatively impact swine reproductive function.     

Epigallocatechin-3-gallate suppresses hemin-aggravated colon carcinogenesis through Nrf2-inhibited mitochondrial reactive oxygen species accumulation

BackgroundPrevious studies have presented evidence to support the significant association between red meat intake and colon cancer, suggesting that heme iron plays a key role in colon carcinogenesis. Epigallocatechin-3-gallate (EGCG), the major constituent of green tea, exhibits anti-oxidative and anti-cancer effects. However, the effect of EGCG on red meat-associated colon carcinogenesis is not well understood.ObjectivesWe aimed to investigate the regulatory effects of hemin and EGCG on colon carcinogenesis and the underlying mechanism of action.MethodsHemin and EGCG were treated in Caco2 cells to perform the water-soluble tetrazolium salt-1 assay, lactate dehydrogenase release assay, reactive oxygen species (ROS) detection assay, real-time quantitative polymerase chain reaction and western blot. We investigated the regulatory effects of hemin and EGCG on an azoxymethane (AOM) and dextran sodium sulfate (DSS)-induced colon carcinogenesis mouse model.ResultsIn Caco2 cells, hemin increased cell proliferation and the expression of cell cycle regulatory proteins, and ROS levels. EGCG suppressed hemin-induced cell proliferation and cell cycle regulatory protein expression as well as mitochondrial ROS accumulation. Hemin increased nuclear factor erythroid-2-related factor 2 (Nrf2) expression, but decreased Keap1 expression. EGCG enhanced hemin-induced Nrf2 and antioxidant gene expression. Nrf2 inhibitor reversed EGCG reduced cell proliferation and cell cycle regulatory protein expression. In AOM/DSS mice, hemin treatment induced hyperplastic changes in colon tissues, inhibited by EGCG supplementation. EGCG reduced the hemin-induced numbers of total aberrant crypts and malondialdehyde concentration in the AOM/DSS model.ConclusionsWe demonstrated that EGCG reduced hemin-induced proliferation and colon carcinogenesis through Nrf2-inhibited mitochondrial ROS accumulation.     

Bisphenol A disrupts granulosa cell function

Because of its widespread use and potential adverse biological effects, bisphenol A (BPA) represents one of the most studied endocrine-disrupting compounds. Within the reproductive system, ovarian granulosa cells have been documented as a target of BPA action, but no consensus has been reached about functional modifications induced by BPA. On these bases, we studied the potential disrupting effects of BPA on the main granulosa cell functional activities, also taking into account a potential interference with the ovarian angiogenic process. Ovarian granulosa cells were isolated from porcine follicles and cultured in the presence or absence of BPA at different concentrations for 48 h. Cell proliferation was studied by measuring adenosine triphosphate content. Progesterone (P4) and estradiol 17β (E2) production was determined by radioimmunoassay. Vascular endothelial growth factor (VEGF) output was quantified by an enzyme-linked immunosorbent assay. Redox status was monitored by measuring superoxide anion and hydrogen peroxide, and by determining the activities of the scavenging enzymes superoxide dismutase, catalase, and peroxidase by colorimetric methods. Granulosa cell proliferation as well as redox status resulted unaffected by BPA. Concentrations of E2 were stimulated by the lower BPA concentration, whereas they were inhibited by the larger doses tested. P4 output was decreased by all BPA concentrations. To the contrary, VEGF production was stimulated. Data indicate that BPA can interfere with reproductive activity by affecting granulosa cell steroidogenesis in vitro; furthermore, BPA can exert a promoting effect on the ovarian angiogenic process by increasing VEGF output in pigs. A disruption of this finely tuned process seems particularly relevant because of the risk of uncontrolled neovascularization.     

Epigallocatechin-3-gallate exerts protective effects against heat stress through modulating stress-responsive transcription factors in poultry

1. The aim of the study was to describe the effect of Epigallocatechin-3-gallate (EGCG), a polyphenol derived from green tea, on activator protein-1 (AP-1) components (phospho-c-Jun and c-Fos), cyclooxygenase-2 (COX-2) and heat shock proteins (HSPs) in the liver of heat-stressed quails.

2. A total of 180 5-week-old female Japanese quails were reared either at 22°C for 24 h/d (thermoneutral, TN) or 34°C for 8 h/d (heat stress, HS) for 12 weeks. Birds in both environments were randomly given 1 of 3 diets: basal diet and basal diet with 200 or 400 mg of EGCG added per kilogram of diet.

3. The hepatic c-Jun, c-Fos, COX-2 and HSPs gene expression for quails reared under the HS environment was greater than those reared under the TN environment. Supplemental EGCG decreased hepatic expression of these proteins at a greater extent under HS than TN.

4. In conclusion, suppression of AP-1 COX-2 and HSPs may partly account for the inhibitory effect of EGCG in heat-stressed quail.     

超微绿茶粉对良凤花鸡免疫机能的影响

为了研究不同添加量的超微绿茶粉对良凤花鸡不同生长阶段免疫功能的影响,试验选用1日龄良凤花鸡商品代肉用仔鸡420只,公母各半,随机分为7个处理组,每个处理设3个重复,每个重复20只,自由饮水、自由采食。试验预试期为3 d,从第13天开始每隔10 d逐只称重,在第34天和第74天从每组内随机选取试验鸡6只(公母各3只),翅静脉采血,测定血液学指标、抗氧化指标、免疫学指标。结果表明:添加一定剂量的绿茶粉可以明显促进肉鸡胸腺的生长(P<0.01),对胸腺指数也有升高作用(P<0.05),极显著提高血清H5型和H9型禽流感抗体效价(P<0.01),极显著提高血清新城疫抗体效价(P<0.01),显著升高1～33 d内白细胞数(P<0.05)和淋巴细胞比例(P<0.01)。说明从生长后期(34～73 d)开始添加1.0%超微绿茶粉不仅能在不影响肉仔鸡生长性能的前提下提高肉仔鸡的免疫机能,同时还减少添加量,降低饲养成本。     

Surgical removal of a granulosa cell tumor from a heifer

A twelve month old, noncycling Holstein-Friesian heifer, was examined because of abnormal udder development. Rectal palpation revealed a tense right uterine horn. A solid mass in the right abdomen was detected by ballottement. A granulosa cell tumor of the right ovary was removed and nine months later the animal conceived.     

Preincubation with green tea polyphenol extract is beneficial for attenuating sperm injury caused by freezing‐thawing in swine

Polyphenols (PFs) extracted from green tea, known to be potent anti‐oxidants, have been reported to be effective in increasing the motility and viability of mammalian sperm, preserved in a liquid form. Therefore, we tested whether PFs might also be effective for maintaining the integrity of frozen‐thawed boar spermatozoa. Ejaculates, collected from Clawn miniature pigs, were diluted in a semen extender containing various amounts of PFs (0, 0.01, 0.05, 0.1 and 0.2% w/v) and then stored at 15°C overnight. The semen samples were processed, using the straw freezing procedure, and then frozen in liquid nitrogen. After rapid thawing at 40°C, the spermatozoa were subjected to several assays to evaluate semen quality. Spermatozoa frozen in a medium containing 0.01% w/v PFs exhibited significantly (P < 0.05) higher degrees of post‐thawed viability and acrosomal integrity than those stored in the absence of PFs. However, no change in the mitochondrial activity was noted between the two groups. The inclusion of 0.01% PFs in the semen extender was significantly (P < 0.05) effective in increasing both the rates of monospermic oocyte formation and of blastocyst formation. These findings indicate that preincubation with the semen extender, containing 0.01% PFs prior to freezing, exerts a protective effect on boar sperm by preventing injuries associated with freezing‐thawing.     

BMP-2 and -6 modulate porcine theca cell function alone and co-cultured with granulosa cells

Bone morphogenetic proteins (BMPs) are emerging as a family of proteins crucial in the regulation of fertility and ovulation rate. We have shown that porcine theca cells express BMP receptors, however, there is a paucity of information regarding the effect(s) of BMPs on theca cell function. The purpose of this study was to investigate the effects of BMP-2 and -6 on theca cells cultured under serum-free conditions in terms of steroidogenesis, cAMP release and proliferation. The study was further extended to determine whether BMP responses in theca cells are affected by the addition of granulosa cells to the culture system. Both BMPs suppressed progesterone and androstenedione synthesis by theca cells (P < 0.05) after 144 h in culture. Oestradiol synthesis was suppressed (P < 0.05) by BMP-2, but not BMP-6, and theca cell proliferation was stimulated (P < 0.05) by BMP-6, but not BMP-2, after 144 h in culture. Both BMP-2 and -6 inhibited cAMP release (P < 0.05) by theca cells. Furthermore, progesterone and androstenedione synthesis by co-cultured theca and granulosa cells were suppressed (P < 0.05) whereas cell proliferation was stimulated (P < 0.05). These results provide strong evidence for a functional BMP system in the porcine ovary and that theca cells are responsive to BMPs in terms of steroidogenesis and proliferation. BMP-2 and -6 may have a role as luteinisation inhibitors in this polyovular species.     

3株猪源绿色气球菌的分离鉴定

为确定云南省不同地区3个猪场仔猪发生非创伤性四肢运动障碍的病原因素,无菌采集病死仔猪深部组织样品,采用血琼脂分离培养病原菌,随后对分离菌株做形态观察、生化试验、药敏试验及16 S rRNA基因测序分析.结果:从3个猪场病料中均分离到菌落及个体形态一致的细菌,其16 S rRNA基因序列与NCBI中7株绿色气球菌参考菌株...     

In vitro culture of bovine embryos in murine ES cell conditioned media negatively affects expression of pluripotency-related markers OCT4, SOX2 and SSEA1

Despite extensive efforts, establishment of bovine embryonic stem (ES) cell lines has not been successful. We hypothesized that culture conditions for in vitro-produced (IVP) embryos, the most used source of inner cell mass (ICM) to obtain ES cells, might affect their undifferentiated state. Therefore, the aim of this work was to improve pluripotency of IVP blastocysts to produce suitable ICM for further culturing. We tested KSR and foetal calf serum (FCS) supplements in SOF medium and ES cell conditioned medium (CM) on IVC (groups: KSR, KSR CM, FCS and FCS CM). Cleavage and blastocyst rates were similar between all groups. Also, embryonic quality, assessed by apoptosis rates (TUNEL assay), total cell number and ICM percentage did not differ between experimental groups. However, expression of pluripotency-related markers was affected. We detected down-regulation of OCT3/4, SOX2 and SSEA1 in ICM of FCS CM blastocysts (p < 0.05). SOX2 gene expression revealed lower levels (p < 0.05) on KSR CM blastocysts and a remarkable variation in SOX2 mRNA levels on FCS-supplemented blastocysts. In conclusion, pluripotency-related markers tend to decrease after supplementation with ES cell CM, suggesting different mechanisms regulating mouse and bovine pluripotency. KSR supplementation did not differ from FCS, but FCS replacement by KSR may produce blastocysts with stable SOX2 gene expression levels.