核酸疫苗研究进展(下)

Sato等报道，在质粒载体上引入大肠杆菌Amp^ 抗性基因(内含有6核苷酸片段的ISS免疫增强序列)，真皮免疫小鼠后ISS增强CTL和Th1细胞的免疫反应。有人提出利用含有病毒免疫原性基因的基因大片段进行基因免疫，以弥补单基因免疫活性的不足。将协同因子如IL-2、IL-12、TCA等的表达质粒与抗原表达质粒混合注射可使Th1型细胞免疫增强，而GMcsf能明显加强体液免疫，     

UHPLC-Q/TOF MS结合HPLC-DAD定性和定量分析硫酸黏菌素可溶性粉中的未知添加物

定性和定量分析一批兽药硫酸黏菌素可溶性粉中的未知添加物。照《中国兽药典》2010年版一部对该批检品用微生物检定法进行含量测定时,发现该样品的抑菌圈为虚圈,用薄层色谱鉴别该样品,未显示与标准品溶液一致的主斑点,怀疑该样品中有处方外非法添加物。采用超高效液相色谱-四级杆-飞行时间质谱(UHPLC-Q/TOF MS)对该样品进行筛查,发现疑似添加物,并使用液相色谱-二极管阵列检测(HPLC-DAD)法进行了双重确证和含量测定。该样品中非法添加物确证为磺胺氯达嗪和甲氧苄啶,添加量分别为58.5 mg/g和13.4 mg/g。本研究通过建立筛查方法为监管部门提供技术支撑,通过分析非法添加物的可能原因为打击兽药处方外非法添加提供了思路。     

Guanidinoacetic acid supplementation in broiler chickens fed on corn-soybean diets affects performance in the finisher period and energy metabolites in breast muscle independent of diet nutrient density

(1) Guanidinoacetic acid (GAA) is the single immediate endogenous precursor of creatine (Cr). It was hypothesised that dietary GAA would have different effects on performance and energy metabolites in breast muscle depending on the nutrient density (ND) of corn-soybean-based diets.

(2) A total of 540 one-day-old male Ross 308 broilers were allocated to 9 dietary treatments with 6 replicates (10 birds each) in a 3 × 3 factorial arrangement with three levels of ND (low, 2800; medium, 2950 and high, 3100 kcal metabolizable energy (ME)/kg; and with the other nutrients being constant relative to ME) and supplemented with three levels of GAA (0, 0.6 and 1.2 g/kg) in a 42-d feeding trial.

(3) In the starter and grower periods, increasing levels of ND improved body weight (BW), average daily gain (ADG), average daily feed intake (ADFI) and feed conversion ratio (FCR), with the exception of ADFI in the starter period. GAA supplementation did not affect performance characteristics. All performance indicators responded markedly to increasing ND in the finisher period, whereas the highest GAA level reduced ADFI compared to the unsupplemented control (156 vs. 162 g/d) and concomitantly FCR (1.81 vs. 1.93). No interactive effects were noted for any performance trait. The high ND diet resulted in more breast meat yield on d42, associated with higher fat content and darker colour compared to the other ND levels. The GAA supplementation did not affect carcass and breast traits. At the end of the experiment, Cr was elevated when feeding GAA at 1.2 g/kg (5455 vs. 4338 mg/kg fresh muscle).

(4) To conclude, ND had a substantial effect on performance and carcass traits, whereas any effect of GAA was limited to FCR in the finisher period and independent of diet ND level.     

Inclusion levels and modes of whole grain incorporation into wheat-based rations differentially influence the performance of broiler chickens

1. The objective was to compare three whole grain (WG) inclusion levels (7.5, 15 and 30%) offered to broiler chickens by three modes of WG incorporation: (i) pre-pellet WG inclusion, (ii) post-pellet WG inclusion as a blend of WG and pelleted concentrate and (iii) post-pellet WG inclusion where WG and pelleted concentrate were provided in separate feed trays against a ground-grain, wheat-based control diet.

2. Ten dietary treatments were offered to 6 replicate cages (6 birds per cage) of male Ross 308 chickens from 7 to 28 d post-hatch. The effects of treatment on relative gizzard weights, gizzard contents, pancreatic weights and pH of gizzard digesta were monitored. Parameters of growth performance, nutrient utilisation (apparent metabolisable energy [AME], metabolisable to gross energy [ME:GE] ratios, nitrogen [N] retention and N-corrected AME [AMEn]), apparent starch and protein (N) digestibility coefficients and disappearance rates in for small intestinal segments and concentrations of free amino acids in plasma taken from the anterior mesenteric vein were determined.

3. Whole grain feeding (WGF) did not influence weight gain, but 30% post-pellet blended and 15 and 30% post-pellet separated treatments significantly depressed (P < 0.05) feed intakes while the 30% post-pellet separated treatment improved (P < 0.01) feed conversion ratios (FCR). WGF regimes significantly increased relative gizzard weights.

4. Overall, WGF generated profound responses in AME, ME:GE ratios, N retention and AMEn that were highly correlated with relative gizzard weights. In general, WGF improved starch and protein (N) digestibilities and again there were some correlations with these outcomes and relative gizzard weights.

5. Post-pellet WG inclusions where WG and pelleted concentrate were offered separately provided chickens with the opportunity to choice feed. Birds showed a preference for the relatively high-protein pelleted concentrate and at 30% WG, this resulted in an improvement in FCR of 7.69% (1.260 versus 1.365; P < 0.001) relative to the ground-grain control diet.     

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.     

Evaluation of vegetable protein in canine diets: Assessment of performance and apparent ileal amino acid digestibility using a broiler model

Recent technological advances in the human food industry with respect to meat processing have decreased the availability of animal proteins to the pet food industry which typically formulates diets with an excess of animal protein. In the long term, this is not sustainable; thus, alternative protein sources need to be investigated. This study examined three canine diets, comparing a typical animal protein‐based diet (control) with two experimental diets where the animal protein was substituted in part with vegetable protein (formulated based either on total protein or amino acid content) using a broiler model. Each diet was fed to six cages each containing two birds from day 15, 18 cages in total (36 birds). Excreta were collected from days 19 to 21. On day 23, birds were euthanized and weighed, and their ileal digesta were collected and pooled for each cage. In addition, one leg per cage was collected for evaluation of muscle mass. Results showed no significant difference in animal performance (feed intake or live weight gain) or muscle to leg proportion across the diets. Birds fed the control diet and the diet balanced for amino acid content exhibited the greatest coefficients of apparent metabolizability for nitrogen (p < .001). Birds fed the diets that contained partial replacement of animal with vegetable protein generally had greater ileal digestibility of amino acids compared to birds fed the control (animal protein) diet. Analysis of excreta showed no dietary difference in terms of dry matter content; however, birds fed the diet balanced for total protein and the diet balanced for amino acid content had significantly greater excreta nitrogen than the control (p = .038). Overall, the study suggests vegetable proteins when formulated based on amino acid content are a viable alternative to animal proteins in canine diets.     

Hyperglycaemia in transition dairy cows: Effects of lactational stage and conjugated linoleic acid supplementation on glucose metabolism and turnover

Supplementing conjugated linoleic acid ( CLA ) is supposed to spare glucose due to the milk fat‐depressing effect of the trans ‐10, cis ‐12 CLA isomer, and allows repartitioning nutrients despite an energy deficiency in early lactation. However, there is still a lack of knowledge in terms of the dynamic pattern of the glucose turnover in transition dairy cows. We hypothesized that dairy cows supplemented with CLA have an altered rate of glucose turnover and insulin sensitivity during early lactation. We conducted three consecutive hyperglycaemic clamps (HGC ) in weeks ?2, +2 and +4 relative to parturition in Holstein cows supplemented daily either with 70 g of lipid‐encapsulated CLA (6.8 g trans ‐10, cis ‐12 and 6.6 g of the cis ‐9, trans ‐11 CLA isomer; CLA ; n  = 11) or with 56 g of control fat ( CON ; n  = 11). From week ?3 up to week +4 relative to parturition, milk yield and dry matter intake (DMI ) were recorded daily, while body weight (BW ) and milk composition were obtained once weekly. Blood samples were taken once weekly and every 30 min during the HGC . Plasma was analysed for concentrations of glucose, fatty acids (FFA ), beta‐hydroxybutyrate (BHB ), insulin, triglycerides and cholesterol. The CLA supplementation did not affect performance and metabolic parameters except for BHB and cholesterol. Furthermore, insulin concentrations and insulin sensitivity were affected by treatment. During the HGC in early lactation, insulin response was lower and decrease in FFA and BHB greater compared with the HGC in week ?2 although glucose target concentration achieved during the steady‐state period was similar for all three HGC . Our findings in terms of insulin and cholesterol suggest that body reserves are preserved through CLA feeding without restraining animal's performance. Furthermore, CLA effects on cholesterol and triglyceride concentrations indicated beneficial effects on hepatic lipid export contributing to an improved efficiency of prevailing metabolites in circulation.