绵羊皮肤中GHR、IGF-1和IGF-1R基因表达的发育性变化及品种特点

采用相对定量反转录多聚酶链式反应 (RT-PCR)方法, 以18S rRNA作内标, 研究了罗米丽(Romilly Hillys)×中国美利奴(新疆军垦型)杂交一代优质细毛羊和哈萨克粗毛羊皮肤中生长激素受体(GHR)、胰岛素样生长因子1(IGF-1)和胰岛素样生长因子1受体(IGF-1R) mRNA发育性变化并进行了品种间比较。分别于30、60、90、135、180和255日龄称重、采毛样, 并于30、90、135和255日龄采皮样。结果表明: 粗毛羊和细毛羊体重、羊毛生长的发育模式没有明显的差异, 30~135日龄体重迅速增加, 135~255日龄增重十分缓慢; 30~135日龄羊毛日增长逐渐增加, 135~180日龄羊毛生长十分缓慢, 而180~255日龄又上升到较高水平。粗毛羊皮肤中GHR mRNA在30~90日龄显著增加 (P<0.05), 90日龄达到高峰, 此后显著下降(P<0.05); 细毛羊在135日龄时GHR mRNA极显著地升高(P<0.01), 此后又极显著地下降。粗毛羊皮肤中IGF-1、IGF-1R mRNA 30~90日龄上升, 90日龄之后极显著下降(P<0.01); 细毛羊皮肤中IGF-1、IGF-1R mRNA出生时较高, 然后逐渐下降。品种之间比较, 细毛羊GHR mRNA出现高峰晚于粗毛羊, 135日龄高峰时显著地高于粗毛羊; 粗毛羊IGF-1、IGF-1R mRNA在90日龄出现高峰, 并极显著或显著地高于细毛羊; 粗毛羊90日龄前GHR、IGF-1和IGF-1R mRNA高于细毛羊, 之后低于细毛羊。结果提示: 绵羊皮肤中GHR、IGF-1和IGF-1R基因表达有特定的发育模式, 并存在品种差异。     

抑制CYP4A11基因表达的siRNAs优选及对血管收缩相关因子的影响

为了考察人细胞中细胞色素P450 4A11(CYP4A11)表达受抑制之后是否会影响血管相关收缩因子的表达,在线设计多个抑制细胞中代谢酶编码基因CYP4A11表达的siRNAs并通过BLAST验证;选择效果好的3对siRNAs合成并转染至EA.hy926细胞株,通过荧光染色和定量多聚酶链反应(quantitative polumerase chain reaction,qPCR)初步检测后,再采用表达量相对更高的MG63细胞系进行验证,发现25 nmol/L的siRNA2抑制效果最好;随后使用25 nmol/L siRNA2抑制CYP4A11在EA.hy926细胞株中的表达,并通过qPCR与Western-blot检测血管收缩相关因子的表达情况,发现促血管收缩相关因子内皮素1受体(endothelin 1 receptor,ET1R)、血管紧张素1型受体(angiotensin type 1 receptor,AT1R)表达下调(P0.05),而内皮型一氧化氮合酶(endothelial nitric oxide synthase,eNOS)的表达显著上调(P0.001)。因此,siRNA2能通过显著抑制EA.hy926细胞中CYP4A11的表达,调节血管紧张的相关指标,有促血管松弛作用的趋势。     

半胱胺盐酸盐和LHRH-A对黄鳍鲷IGF-Ⅰ基因表达和生长的影响

本文以黄鳍鲷 (Sparuslatus)为研究对象 ,利用GeneRaceTM 技术 ,从其肝组织中克隆出类胰岛素生长因子 (IGF Ⅰ )cDNA ,并应用半定量RT PCR方法研究了半胱胺盐酸盐 (Cysteaminehydrochloride)和LHRH A对其肝组织IGF Ⅰ基因表达的影响。黄鳍鲷IGF ⅠcDNA全长为 84 0bp ,编码 185aa多肽 ;序列分析表明 ,黄鳍鲷IGF Ⅰ基因编码的氨基酸序列与金鱼的同源性为 75 8% ,与牙鲆的同源性为 86 5 % ,与同属鲷科的黑鲷同源性高达 10 0 % ,证明鱼类类胰岛素生长因子是非常保守的 ,E区域分析结果表明黄鳍鲷IGF Ⅰ属Ea 4型。在饲料中投喂CSH、LHRH A等添加剂 ,实验组黄鳍鲷鱼种的相对生长率、垂体GH含量、肝脏IGF ⅠmRNA水平均显著高于对照组。以上结果提示 :CSH、LHRH A能促进黄鳍鲷生长激素的合成和IGF Ⅰ基因的表达 ,从而促进鱼种的生长     

The effect of feed restriction on expression of hepatic lipogenic genes in broiler chickens and the function of SREBP1

To study the role of sterol regulatory element-binding proteins (SREBP) in lipogenesis and cholesterol synthesis in the chicken, two experiments were carried out. In the first study, seven-week-old broilers (n = 16) were allocated into 2 groups, fasted for 24 h or refed for 5 h after a 24 h fasting. The mRNA concentrations for SREBPs and other lipogenic genes in the liver were determined by quantitative real time PCR. The hepatic mRNA relative abundance of lipogenic genes and genes involved in cholesterol synthesis were significantly greater (p < 0.001) in the refed broilers. Similar results were demonstrated with Northern analysis. The data suggest that in the liver of fasted broilers, genes associated with lipogenesis and cholesterol biosynthesis were inhibited. Indeed, the mRNA concentrations for fatty acid synthase (FAS), malic enzyme, and stearoyl coenzyme A desaturase were almost undetectable after the 24 h fasting. The data also demonstrated that the expression of lipogenic genes coordinate well as a group during the refeeding period. Second, three small interfering RNA (siRNA) oligonucleotides against SREBP1 were designed to be used in transfecting a chicken hepatocarcinoma cell line LMH. One of the three siRNAs effectively reduced SREBP1 mRNA concentration (p < 0.01). The acetyl coenzyme A carboxylase_α (ACC_α) mRNA was also significantly reduced by the SREBP1 siRNA treatment, suggesting that SREBP1 can upregulate the expression of this lipogenic gene. This siRNA, however, did not affect the mRNA for FAS. Taken together, the RNA interference study showed that SREBP1 has the ability to regulate the expression of ACC_α. This study has helped us understand more about the function of SREBP1 and the physiology of the broiler chickens.     

Influence of early qualitative feed restriction and environmental temperature on long bone development of broiler chickens

This investigation was carried out to study the influence of early qualitative feed restriction and environmental rearing temperature on long bone development in broiler. Energy and protein restriction reduced femur width and humerus weight, but did not affect tibia parameters. Broilers kept at cold environmental temperature showed reduced femur, tibia and humerus length and tibia weight, but the calculated density was not affected by rearing temperature. These findings suggest that qualitative feed restriction and environmental temperature influenced the normal long bone growth; however, bone weight/bone length index (calculated density) was not affected by rearing temperature.     

Improved arterial oxygenation with feed restriction in rapidly growing broiler chickens.

1. Rapidly growing broiler chickens fed ad lib. until 56 days, but feed restricted until 60 days of age, had higher arterial oxygen saturations, lower respiratory frequencies, total ventilations that were not different, and higher tidal volumes compared to those fully fed for 56 days. 2. Arterial oxygen saturation correlated negatively with respiratory frequency, but was not related to total ventilation or tidal volume. 3. Hypoventilation appeared not to be the cause of arterial oxygen desaturation. 4. Arterial oxygen desaturation correlated with the degree of right ventricular hypertrophy.     

Effect of early feed restriction in male broiler chicks on plasma metabolic hormones during feed restriction and accelerated growth

1. Plasma GH was greater (P less than 0.05) on day 12 in ad libitum-fed birds compared to restricted chicks. Conversely, maximum GH levels were found to occur in the nutrient restricted chicks during the period of accelerated growth (day 42). 2. A significant decline in circulating insulin concentrations with advancing age was evident in both ad libitum-fed and restricted chicks. 3. Feed restriction significantly suppressed circulating T3 in restricted chicks, with concentrations returning to control levels upon refeeding. 4. A significant increase in T4 with advancing age was evident in both treatment groups, with T4 being significantly greater in controls compared to restricted chicks at 54 days of age.     

Effect of early feed restriction on physiological responses,performance and ascites incidence in broiler chickens raised in normal or cold environment

Intensive selection of broilers for faster growth and better feed efficiency resulted in greater susceptibility to metabolic disorders such as ascites syndrome, which is one of the major causes of mortality and economic loss in broiler industry. Whereas cool temperature is one of the primary triggers for ascites, early feed restriction (FDR) significantly alleviates its incidence and mortality. However, little is known about effects of FDR, cold environmental temperature and their interaction on physiological responses in broiler chickens. For this purpose, 320 one-day-old male broilers were divided into two treatment groups of Ad libitum (Ad) and feed restricted (FR) with eight pen replicates each. Chickens in FR group underwent feed access limitation from days 7 to 14 of age. On day 21 half of the birds (four pens) in each group exposed to the cold temperature (CT) and the other half (four pens) continued at normal temperature (NT). Average daily feed intake, average daily weight gain and feed conversion ratio (FCR) were measured at days 7, 14, 21, 28 and 42. At 39 and 46 days of age two chicks with a BW around the pen average were selected from each pen and slaughtered after collecting blood samples. Then, relative weight of internal organs and right ventricle weight per total ventricle weight (RV : TV) ratio were calculated. Compared with NT group, CT birds had higher daily feed intake and FCR (P<0.05) from day 28 to 42. Cumulative ascites mortality in CT chickens was higher (P<0.001) than NT chicks. Within the CT group, ascites mortality in FR chickens was reduced (P<0.001) to 1.25% compared with 8.75% in Ad chicks. Birds in CT group had significantly (P<0.05) thicker right ventricle and greater relative weight of heart, hematocrit and triiodothyronine concentration. However, none of these parameters were affected by FDR. Under cold stress conditions, FDR reduced activity of alanine aminotransferase and aspartate aminotransferase (P<0.05). Serum triglyceride, cholesterol, high-density lipoprotein and total protein were not influenced by either temperature or feeding regimen. In conclusion, these findings suggest that FDR reduces ascites incidence mainly by allowing better development of internal organs, which helps them to cope with the high metabolic pressure and suffer less damage.     

Effect of chronic heat stress on gastrointestinal histology and expression of feed intake-regulatory hormones in broiler chickens

Heat stress (HS) dramatically impairs the growth performance of broiler chickens, mainly as a consequence of reduced feed intake due to the loss of appetite. This study was aimed at evaluating the alterations induced by chronic HS conditions on the morphological and morphometric features of the gastrointestinal (GI) tract and on the expression of some enteroendocrine cells (EECs) involved in the regulation of feed intake in chickens. Three hundred male chickens (Ross 308) were divided into two experimental groups and raised either in thermoneutral environment for the whole fattening period (0–41 days) (TNT group) or subjected to chronic HS conditions (30 °C for 24 h/day) from 35 to 41 days (HS group). Samples of proventriculus, duodenum, jejunum and cecum were collected from 24 broilers (12/group). Haematoxylin-eosin was used for the morphometric evaluations, while immunohistochemistry was applied for the evaluation of EECs expressing ghrelin (GHR), cholecystokinin (CCK), neuropeptide Y (NPY), glucagon-like peptide-1 (GLP-1), and serotonin (5-HT). In the proventriculus, HS reduced total wall thickness and mucous layer height (P ≤ 0.01) as well as mean diameter, circumference, and area of the compound tubular glands (P ≤ 0.001) with respect to TNT. The small intestine of HS birds was characterised by decreased villous height and total thickness (duodenum, P ≤ 0.01; jejunum, P ≤ 0.001), whereas crypt depth and width were reduced only in the jejunum (P ≤ 0.01). HS had negligible effects on the morphological aspects of the cecum. In the proventriculus, an increase in GHR and NPY EECs was observed in response to HS (P ≤ 0.001). Similarly, the small intestine villi of the HS group showed greater GLP-1 (P ≤ 0.05), 5-HT (P ≤ 0.001) and CCK (P ≤ 0.01) EECs. Moreover, the expression of 5-HT EECs was higher in the duodenal (P ≤ 0.01) and jejunal (P ≤ 0.01) crypts of HS birds, whereas GLP-1 and CCK EECs increased only in jejunal crypts (P ≤ 0.05). Finally, 5-HT EEC expression was increased in the cecum of HS group (P ≤ 0.01). In conclusion, these outcomes demonstrate that chronic HS induces morphometric alterations not only in the small intestine but also in a key organ such as the proventriculus. Furthermore, HS conditions affect the presence and distribution of EECs, suggesting that some GI peptides and biogenic amine may be implicated in the regulation of appetite and voluntary feed intake in heat-stressed broiler chickens.     

The effect of humidity on growth and feed conversion of broiler chickens

The optimum range of relative humidity for chicken after brooding is between 50 and 70%. The range is between 60% and 80% relative humidity during the brooding period. The higher humidities seem to favor better growth and feed conversion. However, one must be careful since the higher humidities seem to favor problems with wet litter, housing and some diseases. Dehydration did not occur under conditions of these tests. Feed conversion, feathering,pigmentation and gain of the chicken is aversely affected by high relative humidities above 26.7°C temperature. Chicken raised at 35° to 37.8°C regardless of humidity do not perform satisfactorily. The importance of water cannot be overlooked as was shown in these trials.The chicken requires approximately 0.68 kg of water per kg of feed consumed and almost 1.59 kg of water for each kg of gain. If the water is unpalatable, contaminated or high in mineral salts consumption can be reduced thus causing a reduction in growth and efficiency.

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Zusammenfassung Der optimale Bereich der Relativen Feuchte für Hühner nach dem Brüten liegt zwischen 50 und 70%, der Bereich während der Brutperiode zwischen 60 und 80%. Höhere Feuchten scheinen ein besseres Wachstum und eine bessere Nahrungsverwertung zu begünstigen. Jedoch muss man vorsichtig sein, weil die höheren Feuchten Schwierigkeiten wie feuchte Streu und Behausung sowie einige Krankheiten begünstigen. Austrocknung kam unter diesen Versuchsbedingungen nicht vor. Futterverwertung, Gefieder, Pigmentierung und Gewichtszunahme werden durch hohe Relative Feuchten bei Temperaturen über 26,7°C ungünstig beeinflusst. Hühner, die bei 35 bis 37,8°C ohne Rücksicht auf die Relative Feuchte gezüchtet wurden,befriedigten nicht. Die Bedeutung des Wassers darf nicht übersehen werden. Das Huhn benötigt annähernd 0,68 kg Wasser je Kg Futterverbrauch und fast 1,59 kg Wasser für jedes kg Gewichtszunahme. Wenn das Wasser nicht schmackhaft oder verunreinigt ist oder einen hohen Gehalt von Mineralsalzen besitzt, kann der Verbrauch reduziert und so eine Beeinträchtigung des Wachstums und der Leistungsfähigkeit verursacht werden.

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Resume L'humidité optimum pour le développement des poulets fraîchement éclos est située entre 50 et 70%. Durant la couvaison, cet optiumum se situe entre 60 et 80%. Une humidité plus élevée semble favoriser la croissance et une meilleure mise en valeur de la nourriture. Il faut pourtant être prudent dans le réglage de l'humidité, car, si elle est trop élevée, elle semble favoriser certaines maladies et apporte des difficultés d'élevage dues au poulailler lui-même et au fait que la litière est humide. On n'as pas constaté de phénomènes de déshydratation au cours de ces essais. Au-dessus de 26,7°C, une forte humidité influence de façon défavorable la nutrition, le plumage, la pigmentation et le développement des poulets. Des poulets élevés entre 35° et 37,8°C sans qu'on tienne compte de l'humidité ne donnent pas satisfaction. Dans les présents essais, on a également mis en évidence l'importance de l'eau. Les poulets ont besoin d'environ 0,68 kg d'eau par kg de nourriture et d'à peu près 1,59 kg d'eau par kg de poids. Si l'eau a un goût désagréable, si elle est polluée ou si elle présente une forte teneur en sels minéraux, les poulets en boivent moins et leur développement en est diminué d'autant.

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Presented at the Fourth International Biometeorological Congress, New Brunswick, N.J., 26 August–2 September 1966. Scientific Article Number A-1291. Contribution No. 3832 of the Maryland Agricultural Experiment Station.     

Effects of dietary antibiotics on intestinal microflora in broiler chickens

Changes were examined in the intestinal microflora in broiler chickens fed a diet containing antibiotics to obtain fundamental information on the mechanisms of beneficial effect of the antibiotics upon livestock production. Three antibiotics (colistin, bacitracin, and enramycin) were employed as feed additives. Experiments were conducted with broiler chickens in two ways. In one way dietary antibiotics were fed continually at levels approved for use as feed additives for a long term. In the other they were fed the same antibiotics for a short term. Significant changes in microflora were observed mainly in such bacterial groups as aerobic bacteria and Lactobacillus. In the long term administration, three possible modes of variance in the bacterial flora were postulated: Changes directly related to the antibacterial spectrum of antibiotics. Antagonistic changes related to an ecological balance in the bacterial flora. Changes in quantitative balance of bacteria constituting each bacterial group. The change in the intestinal microflora during administration of the antibiotic diet was expressed as a complex form of these transition modes. In the short term administration, it was demonstrated that the effect of the antibiotic diet lingered even 7 days after administration. This suggests that antibiotics used as feed additives may possibly affect the stability of the intestinal microflora.     

Effects of density on perching behavior of broiler chickens

Exercise through perching has been suggested as a way to reduce the impact of leg problems in broiler chickens. It is possible that higher stocking densities may motivate birds to perch more, and perhaps reduce some of the detrimental effects to broiler health seen with high stocking densities. The goal of this research was to investigate the effects of density (10, 15, and 20birds/m(2)) and perch design on the frequency of perch utilization. Mixed sex broilers were assigned to 36 pens in a four perch treatmentxthree density factorial with three replications for each treatment/density combination in a randomized complete block design. Pens were assigned to one of four perch treatments: control (no perches), horizontal (three horizontal perches), angled (three 10 degrees angled perches), or mixed angle (one horizontal, one 10 degrees angled, and one 20 degrees angled perch), at each of the three densities. The results show that although the frequency of perch use was low, (2.6%+/-0.15), significant preferences were clear. Perching frequency was significantly higher when birds were raised at stocking densities of 15 (2.88+/-0.303) or 20birds/m(2) (2.76+/-0.211), as opposed to 10birds/m(2) (2.10+/-0.209) as indicated by a marginally significant main effect (p<0.0597), and a significant linear effect (P<0.0427). The birds used the horizontal perches the most, followed by 10 degrees angled and mixed angle perch treatments with decreasing frequency. An effect of age was found, in that perching increased during the first 4 weeks, and dropped off significantly at the end of the rearing period for all perch treatments. In general, perches closest to the hallway or outside of the house were used more than perches in the middle of the pen. Preferential use of the highest section of the 10 degrees angled perches was also found. The applied perch treatments or densities did not significantly affect final body weight or feed conversion. Percent mortality due to heat stress showed a significant increase with density, while mortality not caused by heat stress did not change with perch treatment or density.