Development of a semi-selective medium and an immunofluorescence colony-staining procedure for the detection of Clavibacter michiganensis subsp. sepedonicus in cattle manure slurry

Various compounds and basal media were tested for their suitability to create a semi-selective medium for isolation ofClavibacter michiganensis subsp.sepedonicus (Cms) from cattle manure slurry containing c. 10⁸ colony forming units (cfu) per ml.Plating efficiency of Cms in yeast glucose mineral medium (YGM) was 104% compared with yeast peptone glucose medium. Nalidixic acid, polymyxin B sulphate and the experimental disinfectant S-0208 inhibited colony growth of cattle slurry bacteria as compared with Cms in YGM. The optimal concentration of these inhibitors in combination was determined by modified agar diffusion tests and by pour plating in 24-well tissue culture plates. The semi-selective medium YGMI consisted of YGM supplemented with nalidixic acid (2 mg/l), polymyxin B sulphate (30 mg/l) and S-0208 (125 mg/l). Plating efficiency varied for Cms between 50.9 and 69.6%, for cattle slurry bacteria between 1.8 and 2.5% and for saprophytes from potato heel end extracts between 11.5 and 27.4%.Differentiation of Cms colonies from other colonies was based on their small and bluish colony morphology in pour plates and on immunofluorescence colony-staining (IFC). IFC of a pure culture of micro colonies of Cms in YGM was possible after one day incubation (colonies c. 5 cells). Green background fluorescence in the agar gels was prevented by addition of Tween 20 (0.1%) to the washing buffer and the use of 1% agar gels. IFC of macro colonies of Cms in YGMI, visible with 4x objective magnification, was possible after 4 days. The detection level of the target organism in artificially inoculated cattle slurry in YGMI based on colony morphology varied between 1.4×10³ and 2.3×10⁴ cfu per ml of cattle slurry. Miniaturized plating combined with IFC, using wells in tissue culture plates (=16 mm), proved suitable for detection, but was c. 30 times les sensitive. The recovery of Cms was negatively correlated with the number of saprophytic colonies in the agar plates (R ²=0.74).     

Effect of diet composition and slaughter weight on animal performance, carcass and meat quality, and fatty acid composition in veal calves

The effect of true milk use in the diet of Spanish Brown Swiss male calves on animal performance, carcass and meat quality, and fatty acids composition was studied. In experiment 1, the effect of milk intake [ad libitum continuous (ADLIB) feed vs. restricted 0.7 during 75 days followed by ad libitum feed (RESTR)] and slaughter endpoint (225 kg vs. 5 month) were studied. In experiment 2, ad libitum concentrate feeding [grain-fed (GF)] was compared with milk supplementation until slaughter [milk-fed (MF)] in calves slaughtered at 345 kg. As regards to milk intake, carcass weight and degree of fatness were higher in the ADLIB group (P<0.05). The RESTR group revealed a higher percentage of saturated fatty acids (P<0.05). In experiment 2, the MF group exhibited a higher fat percentage (P<0.05), lower press and cooking losses (P<0.05), and higher scores for tenderness and juiciness (P<0.05) than the GF group. The percentage of saturated fatty acids was higher in the MF group (P<0.05). The results suggest that true milk use in veal production could be an advantageous alternative in terms on production costs, animal performance, and carcass and meat quality.     

Induction of an antioxidant enzyme system and other oxidative stress markers associated with compatible and incompatible interactions between chickpea (Cicer arietinum L.) and Fusarium oxysporum f. sp.ciceris

To ascertain if active oxygen species play a role in fusarium wilt of chickpea caused by Fusarium oxysporum f. sp. ciceris, the degree of lipid peroxidation (malondialdehyde formation) and the activity levels of diamine oxidase (DAO), an apoplastic H₂O₂-forming oxidase, and several antioxidant enzymes, namely ascorbate peroxidase (APX), catalase (CAT), glutathione reductase (GR), guaiacol-dependent peroxidase (GPX) and superoxide dismutase (SOD), were determined spectrophotometrically in roots and stems of ‘WR315’ (resistant) and ‘JG62’ (susceptible) chickpea cultivars inoculated with the highly virulent race 5 of the pathogen. Moreover, APX, CAT, GPX and SOD were also analysed in roots and stems by gel electrophoresis and activity staining; and the protein levels of APX and SOD in roots were determined by Western blotting. In roots, infection by the pathogen increased lipid peroxidation and CAT and SOD activities, although such responses occurred earlier in the incompatible compared with the compatible interactions. APX, GPX and GR activities were also increased in infected roots, but only in the compatible interaction. In stems, infection by the pathogen increased lipid peroxidation and APX, CAT, SOD and GPX activities only in the compatible interaction, and DAO activity only in the incompatible one. In general, electrophoregrams agreed with the activity levels determined spectrophotometrically and did not reveal any differences in isoenzyme patterns between cultivars or between infected and non-infected plants. Further, Western blots revealed an increase in the root protein levels of APX in the compatible interaction and in those of SOD in both compatible and incompatible interactions. In conclusion, whereas enhanced DAO activity in stems, and earlier increases in lipid peroxidation and CAT and SOD activities in roots, can be associated with resistance to fusarium wilt in chickpea, the induction of the latter three parameters in roots and stems along with that of APX, GR (only in roots) and GPX (only in stems) activities are rather more associated with the establishment of the compatible interaction.     

东北西部沙地近20年地下水变化动态及其成因分析

本文以奈曼旗为例，分析研究了我国东北西部沙地1979-1997年地下水位时空变化规律及其与降水量和沙漠化的关系。结果表明：（1）沙地多年平均地下水位为2.19m，其中农区为2.85m，牧区1.16m;(2)地下水位是1981-1983年最低为2.49-2.56m,1991-1992年最高为1.90-1.92m，1997年为2.22m，高于1979年的2.41m；（3）该区季节最高水位主要发生于春季（3-5月），发生频率占50.1% ,最低水位主要发生于冬季（12-2月），发生频率为41.2%，平均4月最高为2.07m，10月最低为2.23m;(4)地下水位的年际变化动态和降水量密切相关，但有滞后1-2年的现象；（5）地下水位空间分布上的差异与土地沙漠化及土地利用方式密切相关，沙漠化土地主要用于放牧，由于人为利用少和蒸散少，有利于地下水抬升；（6）由于耕地面积占总土地面积比例低，水浇地和粮食产量的增加尚未对地下水位造成严重影响。     

低温胁迫下弓葵幼苗膜脂过氧化及保护酶活性的变化

 低温胁迫下弓葵( Butia capitata Becc) 幼苗叶片的MDA 含量逐渐增加, 膜脂过氧化作用增强。- 8 ℃条件下的膜脂过氧化作用明显强于2 ℃。细胞膜透性在2 ℃条件下变化不大, - 8 ℃时则随低温胁迫时间延长而急剧上升, 细胞膜受到伤害。- 8 ℃胁迫下细胞保护酶SOD、POD 和CAT 活性短期(6 h) 内升高,然后下降, 24 h 以后3 种保护酶受到低温胁迫的严重抑制。在2 ℃胁迫下, SOD 活性在6 h 内变化不大, 随后下降; CAT活性变化趋势与- 8 ℃时相似, 但变化幅度较小; POD 虽也呈现先升后降的趋势, 但降幅明显小于升幅, 至48 h 时POD 活性仍维持较高水平。2 ℃低温胁迫不是抑制而是促进POD 活性的提高。     

尿素水平对青山羊血液生化指标的影响

应用饲养试验与氮平衡试验,研究了不同尿素水平的日粮对青山羊血清总蛋白(STP)、血清清蛋白(ALB)、血清球蛋白(GLB)、血清清球蛋白比值(A/G)、血清尿素氮(SUN)及血清谷丙转氨酶的含量变化的影响。试验采用4×4拉丁方试验设计,试验分4个阶段,在基础日粮一致的情况下,采用4、9、14、0(对照)g/kg·DM4种不同的处理,试验结果表明:不同尿素水平极显著影响ALB、SUN浓度和GPT活性浓度(P<0.01),对血清清球比(A/G)无显著差异(P>0.05),尿素水平与STP、GLB浓度存在负相关;血清含氮生化指标STP、ALB、GLB、A/G、GPT、SUN随时间呈动态变化,出现高峰的时间比瘤胃氨浓度出现的时间平均推迟1～2h。进食尿素后,血液生化指标由平稳逐渐缓慢升高到高峰,随后又下降并趋于平衡。     

酵母铬与L-肉碱对蛋鸡生产性能及脂质代谢的影响

选用21周龄海兰褐壳蛋鸡288只,随机分为9组,每组32只,采用3×3(铬×L-肉碱)2因子3水平有重复交叉试验设计,预试2周后,在玉米-豆粕型日粮的基础上添加不同水平的酵母铬(以铬计:0、400、600μg/kg)与L-肉碱(0、50、100mg/kg),以饲喂基础日粮组为对照组,用以研究二者对蛋鸡生产性能及脂质代谢的影响,并初步探讨二者的互作效应,试验期7周。结果表明:600μg/kg铬或100mg/kgL-肉碱对脂质代谢多数指标效果显著(P<0.05或0.01);二者同时添加对蛋黄胆固醇、肝脏甘油三酯及腹脂率等均产生显著互作效应(P=0.0003～0.0500),且400μg/kg铬和100mg/kgL-肉碱混合添加对降低蛋黄胆固醇比其它各组均表现出优势,400μg/kg铬和50mg/kgL-肉碱混合添加对降低腹脂率比其它各组均表现出优势。铬或(与)L-肉碱在降低蛋鸡肝脂、腹脂及蛋黄胆固醇的同时并未影响蛋鸡的生产性能。     

开食料中不同NDF水平对犊牛生长性能、瘤胃内环境及血清生化指标的影响

为探索开食料中不同NDF水平对犊牛生长、瘤胃发酵参数以及血清生化指标的影响,选用初生重相近(42±2.5) kg、饲喂足量初乳的中国荷斯坦犊牛60头,其中公犊牛36头,母犊牛24头。随机分为A、B、C和D 4个处理组,每个处理15头,其中9头公犊牛+6头母犊牛,试验犊牛于犊牛岛(1.5 m×3.4 m)单独饲养。15日龄开始分别饲喂NDF水平不同的4种开食料,分别为:A处理10%,B处理15%,C处理20%和D处理25%,70日龄断奶,试验期112 d。结果表明:1)0~42日龄和70~112日龄,B组日增重均高于A、C和D组(P<0.05),分别高出45.69、39.79、117.13 g·d^-1和35.33、153.29、 145.93 g·d^-1。70~112日龄,B、C和D组开食料采食量显著高于A组(P<0.05),分别高出255.20、252.48和392.27 g·d^-1。A(0.61)和B(0.60)组犊牛的饲料转化率显著高于C(0.55)和D(0.54)组(P<0.05);2)犊牛瘤胃pH在35和70日龄时,C组显著高于其他3组(P<0.05),112日龄时,A和B组显著高于C和D组(P<0.05);3)瘤胃NH₃-N含量在90日龄前差异不显著(P>0.05),但在112日龄时, B、C和D组较A组显著降低16.72%、44.19%和52.10%(P<0.05);4)C和D组乙酸比例显著高于A和B组(P<0.05);B组丙酸比例较A、C和D组高出4.24%、3.18%和6.43% (P<0.05),乙酸/丙酸显著低于A、C和D组(P<0.05);5)各处理组间血清总蛋白、葡萄糖和尿素氮含量无显著差异(P>0.05)。1~3月龄犊牛开食料中NDF适宜水平为15%有利于提高断奶后日增重,促进饲料利用效率,改善瘤胃内环境,且对犊牛血清生化指标无不利影响。     

Branched‐chain amino acid ratios in low‐protein diets regulate the free amino acid profile and the expression of hepatic fatty acid metabolism‐related genes in growing pigs

Liver metabolism is affected by nutrients. The aim of this study was to explore the effects of low‐protein diets (17% crude protein, CP) supplemented with branched‐chain amino acids (BCAAs), including leucine (Leu), isoleucine (Ile) and valine (Val), on hepatic amino acid profile and lipid metabolism in growing pigs. The ratio of Leu : Ile : Val in all groups was 1 : 0.51 : 0.63 (20% crude protein, CP), 1 : 1 : 1 (17% CP), 1 : 0.75 : 0.75 (17% CP), 1 : 0.51 : 0.63 (17% CP) and 1 : 0.25 : 0.25 (17% CP) respectively. Results revealed that compared to the positive control group (1 : 0.51 : 0.63, 20% CP), the low‐protein diets significantly augmented the concentrations of most essential amino acids and non‐essential amino acids (p < .05), with the greatest values observed in the 1 : 0.25 : 0.25 group. Moreover, relative to the control, the low‐protein diets with the Leu : Ile : Val ratio ranging from 1 : 0.75 : 0.75 to 1 : 0.25 : 0.25 markedly downregulated the mRNA abundance of acetyl‐CoA carboxylase (ACC), lipoprotein lipase (LPL) and fatty acid‐binding protein 4 (FABP‐4) (p < .05), and upregulated the mRNA expression of hormone‐sensitive lipase (HSL), peroxisome proliferator‐activated receptor‐g coactivator‐1α (PGC‐1α), uncoupling protein 3 (UCP3) and liver carnitine palmitoyltransferase 1 (L‐CPT‐1) (p < .05). Therefore, our data suggest that protein‐restricted diets supplemented with optimal BCAA ratio, that is, 1 : 0.75 : 0.75–1 : 0.25 : 0.25, induce a shift from fatty acid synthesis to fatty acid oxidation in the liver of growing pigs. These effects may be associated with increased mitochondrial biogenesis.