功能基因组学在鉴别新的卵蛋白质中的应用

学角度而言,鸡蛋就像一个天然容器,它能够提供胚胎宫外发育所需的营养物质和生物活性分子[1,2].这意味着,鸡蛋里面必须包含能使生殖细胞发育为成熟小鸡的所有成分.鸡蛋中含有胚胎发育所需的维生素和蛋白质(蛋清和蛋黄),脂肪(蛋黄)和矿物质(蛋壳).鸡蛋也是人类不可缺少的食物.鸡蛋中氨基酸比例均衡有利于胃肠道吸收,因而营养价值很高[3,4].     

Altitude effects on technology and productivity of small bovine farms (milk meat) in Veracruz (Gulf of Mexico)

The dual-purpose bovine system represents 98.4% of the bovine livestock of Veracruz, the main cattle-producing state of Mexico. This system supplies calves to meat companies, a sector in which Veracruz has been the national leader in the last decade. The objective of the present study was to analyze the effect of the altitudinal zonation of farms on livestock technology and productivity in a microbasin of the Gulf of Mexico where small farms predominate. Structured interviews were applied to producers located in three altitudinal zones (at average altitudes of 50, 140, and 450 m, respectively, for lower, middle, and upper zones). Sample size was 135 farms having similar land surface (within a range of 15–22 ha). The results indicated multiple differences among farms located in the three zones. Farms in the middle and lower zones presented higher productive indicators than those in the upper zone. Differences in herd structure and management resulted in important differences in productivity, income, and profits in milk and calf production. We concluded from this study that altitudinal zonation in Veracruz had a clear effect on the differentiation of small farms, which are representative of dual-purpose cattle. The upper zone performs cattle activity under conditions with greater disadvantages in the analyzed region.     

Effects of protein-energetic supplementation frequency on growth performance and nutritional characteristics of grazing beef cattle

Two experiments were conducted to evaluate reduced supplementation frequencies for grazing beef cattle in rainy season. In experiment 1, evaluating the nutritional parameters, four rumen-cannulated Nellore bulls (BW = 410 kg) were used. In experiment 2, evaluating animal performance, 48 Nellore bulls (BW = 358 kg) were used. The treatments were as follows: mineral supplement (MS) alone and MS plus protein-energy supplement provided 3×, 5× and 7×/week. Supplementation frequency did not affect (P > 0.05) intake and digestibility. Average daily gain was greater (P < 0.001) to supplementation compared with MS. The supplementation 5×/week resulted in greater weight gain per hectare (9.24) and higher economic returns during the study period (1.64%) compared to other supplementations. Supplementation 5×/week increased animal performance and positively influenced economic returns.     

The NS3 proteins of global strains of bluetongue virus evolve into regional topotypes through negative (purifying) selection

Comparison of the deduced amino acid sequences of the genes (S10) encoding the NS3 protein of 137 strains of bluetongue virus (BTV) from Africa, the Americas, Asia, Australia and the Mediterranean Basin showed limited variation. Common to all NS3 sequences were potential glycosylation sites at amino acid residues 63 and 150 and a cysteine at residue 137, whereas a cysteine at residue 181 was not conserved. The PPXY and PS/TAP late-domain motifs were conserved in all but three of the viruses. Phylogenetic analyses of these same sequences yielded two principal clades that grouped the viruses irrespective of their serotype or year of isolation (1900-2003). All viruses from Asia and Australia were grouped in one clade, whereas those from the other regions were present in both clades. Each clade segregated into distinct subclades that included viruses from single or multiple regions, and the S10 genes of some field viruses were identical to those of live-attenuated BTV vaccines. There was no evidence of positive selection on the S10 gene as assessed by reconstruction of ancestral codon states on the phylogeny, rather the functional constraints of the NS3 protein are expressed through substantial negative (purifying) selection.     

Evaluation of antibody-ELISA and real-time RT-PCR for the diagnosis and profiling of bluetongue virus serotype 8 during the epidemic in Belgium in 2006

In 2006 bluetongue (BT) emerged for the first time in North-Western Europe. Reliable diagnostic tools are essential in controlling BT but data on the diagnostic sensitivity (Se) and specificity (Sp) are often missing. This paper aims to describe and analyse the results obtained with the diagnostics used in Belgium during the 2006 BT crisis. The diagnosis was based on a combination of antibody detection (competitive ELISA, cELISA) and viral RNA detection by real-time RT-PCR (RT-qPCR). The performance of the cELISA as a diagnostic tool was assessed on field results obtained during the epidemic and previous surveillance campaigns. As the infectious status of the animals is unknown during an epidemic, a Bayesian analysis was performed. Both assays were found to be equally specific (RT-qPCR: 98.5%; cELISA: 98.2%) while the diagnostic sensitivity of the RT-qPCR (99.5%) was superior to that of the cELISA (87.8%). The assumption of RT-qPCR as standard of comparison during the bluetongue virus (BTV) epidemic proved valid based on the results of the Bayesian analysis. A ROC analysis of the cELISA, using RT-qPCR as standard of comparison, showed that the cut-off point with the highest accuracy occurred at a percentage negativity of 66, which is markedly higher than the cut-off proposed by the manufacturer. The analysis of the results was further extended to serological and molecular profiling and the possible use of profiling as a rapid epidemiological marker of the BTV in-field situation was assessed. A comparison of the serological profiles obtained before, during and at the end of the Belgian epidemic clearly showed the existence of an intermediate zone which appears soon after BTV (re)enters the population. The appearance or disappearance of this intermediate zone is correlated with virus circulation and provides valuable information, which would be entirely overlooked if only positive and negative results were considered.     

Identification and validation of housekeeping genes as internal control for gene expression in an intravenous LPS inflammation model in chickens

Real-time PCR has become a powerful tool for the detection of inflammatory parameters, including cytokines. Reference or housekeeping genes are used for the normalization of real-time RT-PCR results. In order to obtain reliable results, the stability of these housekeeping genes needs to be determined. In this study the stability of five genes, including beta-actin, glyceraldehyde-3-phosphate dehydrogenase (GAPDH), hypoxanthine phophoribosyl-transferase (HPRT), ubiquitin (UB) and glucose-6-phosphate dehydrogenase (G6PDH), was determined in a lipopolysaccharide inflammation model in chickens. beta-Actin appeared to be the most stable single gene in our model. Because the use of a single gene for normalization can lead to relatively large errors, the use of the geometric mean of multiple reference genes or normalization factor is preferred. The most stable combination for gene expression analysis in this lipopolysaccharide inflammation model in chickens is G6PDH and UB, since their correlation coefficients were 0.953 and 0.969, respectively (BestKeeper) and an M value of 0.34 and a low V(2/3) value of 0.155 (geNorm) were obtained. The use of HPRT and GAPDH should be avoided. The stable housekeeping genes, G6PDH and UB together, can be used to normalize the expression of pro-inflammatory cytokines in a lipopolysaccharide inflammation model in chickens.     

Cluster of cases of malignant schwannoma in cattle

Between 1998 and 2001, several cases of ataxia and paresis followed by recumbency and death were reported in cows from different farms in a restricted area of the Argentinian Patagonia. Five cases of this cluster were studied and a diagnosis of malignant schwannoma was established. Electron microscopy (em) of tumour samples from three of the animals revealed intracytoplasmic or interstitial structures resembling retroviral particles. Attempts to isolate a viral agent from the tumours were unsuccessful but the epidemiological data and the em findings suggest a viral aetiology.     

营养水平对妊娠早期母猪胚胎存活和瘦素、孕酮分泌及基因表达影响

本试验选用63头长白-大白杂交初产母猪,研究营养水平对初产母猪妊娠早期胚胎存活和瘦素(leptin)、孕酮分泌及胚胎瘦素、瘦素受体(Ob-R)、信号传导和转录活化因子-3(STAT-3)、孕酮受体(PGR)、DNA甲基化转移酶-1(DNMT-1)基因表达的影响.将配种后的母猪随机分到高、中、低3个营养水平组按2倍、1.2倍和0.6倍维持需要(分别记为2.0M、1.2M和0.6M)供给饲粮,妊娠12、25和35 d屠宰母猪收集血清和胚胎,用ELISA方法测定血清中瘦素、孕酮水平,用RT-PCR方法研究目的基因在胚胎中的表达差异.结果表明:①胚胎存活率,妊娠12、25和35 d时,1.2M组均显著高于2.0M组(P<0.05);1.2M组妊娠12 d时与0.6M组间差异不显著(P>0.05),而妊娠25、35 d高于0.6M组(P<0.05);②血清瘦素水平,妊娠12 d时,2.0M组显著高于1.2M组(P<0.05),极显著高于0.6M组(P<0.01);妊娠25和35 d时,2.0M组极显著高于1.2M和0.6M组(P<0.01);1.2M组极显著高于0.6M组(P<0.01).血清孕酮水平,妊娠12、25和35 d时,2.0M组极显著低于1.2M组和0.6M组(P<0.01),1.2M组和0.6M组之间差异不显著(P>0.05);③营养水平对胚胎存活重要基因表达的影响,leptin和ob-R基因mRNA的相对表达量在妊娠12、25和35 d时,2.0M组显著高于1.2M和0.6M组(P<0.05),而1.2M与0.6M组差异不显著(P>0.05).妊娠12和25 d时,2.0M组胚胎STAT-3基因的相对表达极显著高于0.6M组(P<0.01);妊娠35 d时,2.0M和1.2M组极显著高于0.6M组(P<0.01).妊娠12、25和35 d时,2.0M组胚胎DNMT-1的相对表达显著高于0.6M组(P<0.05).妊娠12 d时,各水平组胚胎PGR表达差异不显著(P>0.05),妊娠25 d时,0.6M组极显著高于2.0M组(P<0.01),显著高于1.2M组(P<0.05),妊娠35 d时,1.2M组极显著高于2.0M和0.6M组(P<0.01),0.6M组显著高于2.0M组(P<0.05).以上结果表明在妊娠35 d以内以中营养水平饲喂初产母猪具有较高的胚胎存活率.营养水平显著影响瘦素、孕酮的分泌及lep-tin等基因的表达,瘦素、孕酮分泌过高、过低均对胚胎发育不利,高水平饲喂及严重限饲引起胚胎中leptin、Ob-R、STAT-3、PGR、DNMT-1基因表达变化,降低胚胎存活率.     

A Single Sample Method for Evaluating 51Chromium-Ethylene Diaminic Tetraacetic Acid Clearance in Normal and Hyperthyroid Cats

Background: Chronic kidney failure is frequently seen in middle-aged and elderly cats. ⁵¹Chromium-ethylene diaminic tetraacetic acid (⁵¹Cr-EDTA) clearance and single blood sample (SBS) method are used in several species to estimate the glomerular filtration rate (GFR).
Hypothesis: The hypothesis of this study was that ⁵¹Cr-EDTA clearance could be determined using an SBS method in normal and hyperthyroid cats.
Animals: Forty-six cats were included in this study, with an average age of 9.5 years. Of these cats, 27 had hyperthyroidism; 19 were healthy.
Methods: After IV injection of ⁵¹Cr-EDTA (average dose: 4.25 MBq), 7 blood samples were obtained between 5 and 240 minutes. Reference clearance was calculated in mL/min and mL/min/kg body weight, using a 2-compartment model. Optimal time for clearance measurement with SBS was then determined by systematically comparing each individual plasma concentration to the reference multisample clearance.
Results: The average reference plasma clearance of ⁵¹Cr-EDTA for all cats was 14.9 mL/min (3.7 mL/min/kg). The clearance in hyperthyroid cats averaged 16.4 mL/min (4.3 mL/min/kg) and in normal cats averaged 10.3 mL/min (2.4 mL/min/kg).
The optimal time for the SBS was 48 minutes after injection of tracer ⁵¹Cr-EDTA ( R ²= 0.9414), giving the following converting equation: clearance = (0.0066 × DV_{48 minutes}) – 0.9277 (in mL/min).
Conclusions and Clinical Importance: In this study, the single sample ⁵¹Cr-EDTA clearance method was used to estimate the global GFR in cats. The method identified differences in clearance between normal and hyperthyroid cats. The optimal time for an SBS was 48 minutes.