Molecular Characterization of the Von Willebrand Factor Type D Domain of Vitellogenin from Takifugu flavidus

The von Willebrand factor type D (VWD) domain in vitellogenin has recently been found to bind tetrodotoxin. The way in which this protein domain associates with tetrodotoxin and participates in transporting tetrodotoxin in vivo remains unclear. A cDNA fragment of the vitellogenin gene containing the VWD domain from pufferfish (Takifugu flavidus) (TfVWD) was cloned. Using in silico structural and docking analyses of the predicted protein, we determined that key amino acids (namely, Val¹¹⁵, ASP¹¹⁶, Val¹¹⁷, and Lys¹²²) in TfVWD mediate its binding to tetrodotoxin, which was supported by in vitro surface plasmon resonance analysis. Moreover, incubating recombinant rTfVWD together with tetrodotoxin attenuated its toxicity in vivo, further supporting protein–toxin binding and indicating associated toxicity-neutralizing effects. Finally, the expression profiling of TfVWD across different tissues and developmental stages indicated that its distribution patterns mirrored those of tetrodotoxin, suggesting that TfVWD may be involved in tetrodotoxin transport in pufferfish. For the first time, this study reveals the amino acids that mediate the binding of TfVWD to tetrodotoxin and provides a basis for further exploration of the molecular mechanisms underlying the enrichment and transfer of tetrodotoxin in pufferfish.     

牛源荚膜A型多杀性巴氏杆菌疫苗候选株的筛选及鉴定

为了筛选生长快、毒力强、免疫原性好、副反应小的牛源荚膜A型多杀性巴氏杆菌（Pasteurella multocida，Pm）灭活疫苗菌株，本试验选取6株来自不同地区致犊牛肺炎死亡的牛源荚膜A型多杀性巴氏杆菌分离株，测定了培养基生长曲线、小鼠毒力、菌体脂多糖（LPS）含量及各菌株灭活菌苗免疫小鼠和家兔后的抗体效价，并进行了攻毒保护试验。结果显示，分离株Pm2、Pm3、Pm5生长速度较快、毒力较强、LPS含量较多，均含有与毒力和免疫相关的ptfA和fimA基因；免疫小鼠及家兔未发现明显不良反应，在二免后14 d血清抗体达1：64~1：128，强毒攻毒后全部存活，而PBS对照组全部死亡。本试验结果表明，Pm2、Pm3、Pm5均可作为多杀性巴氏杆菌灭活菌苗的候选菌株，其中Pm3作为首选株。     

Application of molybdenum fertilizer enhanced quality and production of alfalfa in northern China under non-irrigated conditions

Molybdenum (Mo) is an essential micro-mineral nutrient for plant growth and development. As an important legume forage, alfalfa (Medicago sativa) is frequently planted in marginal lands of northern China where water scarcity and infertile soil are limiting factors for alfalfa production. Mo plays an important role in nitrogen (N) metabolism in general and N fixation in legume species in particular. To evaluate the role of Mo fertilizer in alfalfa quality and production in semi-arid areas of northern China, we conducted a three-year field experiment in Inner Mongolia from 2012 to 2014. Mo fertilizer at the doses of 0, 150 g hm^?2, 300 g hm^?2yr^?1 was applied consecutively across the three years. Application of Mo at 300 g hm^?2yr^?1 significantly increased dry yield of alfalfa in 2014. Crude protein (CP) content of alfalfa without application of Mo exhibited a decline trend from 2012 to 2014. Application of Mo significantly enhanced total CP in 2012 and 2014, and delayed the reduction in total CP across the three years. In contrast to CP, Mo fertilizer had no significant effects on neutral detergent fiber and acid detergent fiber content. In addition, we found that nitrate reductase activity of alfalfa was increased by Mo fertilizer, which may account for the enhanced CP content in alfalfa. These findings highlight that application of Mo fertilizer can be an effective way to improve alfalfa production and quality in semi-arid areas in northern China.     

Choline supplementation increased total body lipid gain,while surplus methionine improved growth and amino acid retention in adult Atlantic salmon (Salmo salar)

Methionine–choline‐deficient (MCD) mammals are known to accumulate liver TAG probably due to phosphatidylcholine (PC) deficiency and thus assembly of VLDL and transport of lipids from liver to peripheral organs. To assess whether supplementation of choline could spare methionine and secure a healthy liver metabolism, by reducing the endogenous PC synthesis without interfering with lipid transport and distribution, Atlantic salmon with initial BW of 700 g were fed adequate (1.9 g Met/16 gN) or surplus methionine (2.5 g Met/16 gN) diets of which were supplemented with choline or not for a period of 19 weeks. Fish fed the low‐methionine diets had reduced growth (p = .013) due to reduced protein gain (p = .007), while lipid gain slightly improved in fish fed the choline‐supplemented diets (p = .047). Also, feed conversion improved when fed surplus methionine (p < .001), while choline supplementation had no impact on feed conversion. No interaction between choline and methionine on growth performance or retention existed. Phospholipid status in liver and muscle was not affected by treatments, and no liver TAG accumulation occurred at the methionine levels used. Gene expression of ApoB100 necessary for assembling VLDL or pemt necessary for endogenous PC synthesis was unaffected by treatments. Capacity of methylation (MAT, BHMT) within the liver was not affected by treatment nor was the gene expression of enzymes in liver transsulfuration (CBS or CDO). Methionine status within liver was unaffected by treatments, while free methionine reduced in those fish fed the low‐methionine diets in muscle and plasma. Cystathionine and taurine were elevated when fed surplus methionine. Choline supplementation had no impact on sulphur amino acid metabolites in either tissue. Neither did choline supplementation improve TAG mobilization from liver to muscle. To conclude, choline does not improve endogenous phospholipid synthesis or transport of TAG from liver to muscle depot when added to diets containing 1.9 g Met/16 gN, while surplus methionine improved growth and protein retention, indicating that 1.9 g Met/16 gN is enough to support a healthy liver metabolism, but too low to support muscle protein deposition in adult salmon fed high plant protein diets for longer periods of time.     

Effects of dietary protein to energy ratios on growth,body composition and digestive enzyme activities in Chinese mitten‐handed crab,Eriocheir sinensis

This study aimed to determine the optimal protein to energy ratio (P/E ratio) and evaluate the effect of dietary protein and lipid levels on growth performance, body composition and digestive enzymes activities in Chinese mitten‐handed crab, Eriocheir sinensis. Nine practical diets containing three levels both for protein (DP 30%, 35% and 40%) and lipid (DL 2%, 7% and 12%) with P/E ratios ranging from 13.69 to 19.79 mg KJ^?1 were fed to four replicates of crabs (3.39 ± 0.10 g) for 10 weeks. Weight gain increased significantly with the increase in DP level at each DL level. Moreover, weight gain increased in crabs fed with diets containing DL level from 2% to 12% and DP level from 30% to 35%. However, the diet containing 40% DP and 12% DL levels significantly decreased the growth performance and protein efficiency of the crabs. The whole crab and hepatopancreas lipid contents also increased as dietary lipid increased, but not dietary protein. The total protease activity increased significantly with the increase in dietary protein at each lipid level. The lipase activity was statistically comparable among different DL levels at each DP level. Taken together, the crab fed the diet containing 35% protein and 12% lipid levels with P/E 15.77 mg KJ^?1 revealed optimal growth, feed utilization efficiency and digestive enzymes activities. Moreover, our study indicated that the higher dietary lipid level at a relatively lower dietary protein level could provide protein sparing effect in Eriocheir sinensis.     

Using near‐infrared reflectance spectroscopy to predict the digestible protein and digestible energy values of diets when fed to barramundi,Lates calcarifer

This study examined the potential of using near‐infrared spectroscopy (NIRS) to predict nutrient digestibility parameters (digestible protein and digestible energy) of compound diets when fed to barramundi. A series of 60 diets were assessed for their protein and energy digestibilities in a series of five experiments over a 5‐year period from 2009 to 2014. Considerable variance was observed in the digestibility parameters of diets across the experiments, providing a suitable range in diet digestible protein and digestible energy values from which to develop a NIRS calibration. Samples of the same diets were also scanned using a diode array near‐infrared spectrophotometer (DA‐NIRS). The spectra were obtained by the DA‐NIRS and were chemometrically calibrated against the digestible value data using multivariate analysis software. The results in terms of standard error of cross‐validation (SECV), residual prediction deviation (RPD) and correlation coefficient (R²) show good relationships (R² > 0.8) between the predicted and observed parameters for both the digestible protein and digestible energy parameters assessed. This study therefore demonstrates that it is possible to use NIRS technology to provide rapid estimates of the digestible protein and digestible energy values of compound diets for barramundi in near real time.     

杂草环境下转EPSPS基因大豆NZL06-698的生态适应性研究

为研究转EPSPS基因大豆NZL06-698在杂草环境中的生态适应性,试验设置杂草处理模拟野外环境,研究转基因大豆NZL06-698(GT698)的生存竞争能力以及外源EPSPS蛋白表达情况。结果表明:在相同处理下转基因大豆GT698的株高(R8期)、百粒重显著高于亲本大豆蒙豆12(MD12),但单株产量、单株籽粒数、结实率显著低于亲本大豆,说明外源基因的存在并未增强转基因大豆的生存竞争力,且转基因大豆在野外的生存竞争能力与亲本大豆相比较弱。试验注意到,杂草环境对转EPSPS基因大豆的生长有明显的限制作用,杂草处理下转基因大豆的单株产量、单株籽粒数、百粒重、结实率等指标均显著低于对照处理。ELISA检测结果表明,转基因大豆叶片及籽粒中外源EPSPS蛋白在杂草处理及对照中均正常表达,且两种处理下的EPSPS蛋白表达量无显著差异。以上结果表明杂草环境中转基因大豆的EPSPS蛋白正常表达,正常提供抗草甘膦性状,但是外源基因的存在并没有增加转基因大豆的生存竞争能力,且转基因大豆的生长受到杂草环境的显著抑制,由此得出结论:与亲本大豆相比,转EPSPS基因大豆NZL06-698在野外环境中生态适应能力较弱。     

草坪草屑的加工与利用

我国每年修剪草坪产生大量的草坪草屑,对草坪草屑的加工利用,可以对我国能源、饲料、肥料及蛋白资源的短缺起到一定的缓解作用,因此如何对草坪草屑进行合理利用已成为一项重要的研究课题。本文综述了草坪草屑能源化、饲料化、肥料化和深加工4种利用方式,详细介绍了草坪草屑在发酵生产能源、饲喂家畜、堆肥还坪以及加工叶蛋白等方面的研究进展,指出禾本科草坪草屑和豆科草坪草屑应采用不同的处理方式,对各加工方式的投入和产出也应进行进一步的量化核算,同时总结并展望了草坪草屑利用方式的发展前景。     

Apparent digestibility coefficients of animal and plant feed ingredients for juvenile Pseudobagrus ussuriensis

A study was conducted to determine apparent digestibility coefficients (ADCs) of dry matter (DM), crude protein (CP), lipid, gross energy, phosphorus and amino acids (AAs) from Peruvian fish meal (PFM), native fish meal (NFM), meat bone meal (MBM), dried daphnia meal (DDM), soybean meal (SBM), full‐fat soybean (FSBM), cottonseed meals (CSM), rapeseed meal (RSM) and corn gluten meal (CGM) for Pseudobagrus ussuriensis (7.8 ± 0.06 g). Test diets were formulated to contain a 70:30 mixture of reference diet to test ingredient with chromic oxide (5 g kg^–1) serving as the inert marker. ADCs of DM ranged from 49.94% (CGM) to 87.11% (PFM), the highest ADCs of DM were observed in PFM and the lowest was found in CGM. ADCs of CP in the test ingredients ranged from 74.92% (CGM) to 93.41% (SBM). ADCs of crude lipid ranged from 51.80% (CGM) to 94.07% (PFM) for fish. ADCs of energy ranged from 56.42% (CGM) to 90.09% (PFM). ADCs of phosphorus ranged from 21.81% (CSM) to 65.70% (PFM). AA availability for the test ingredients followed a similar pattern to ADCs of CP, in which values for PFM, NFM and MBM were generally higher (p < .05) than those of other protein ingredients. Among all plant meals, the AA availability values in SBM were higher (p < .05) than those in CGM, CSM and RSM. The availability of most AAs in CGM was the lowest (p < .05) among all tested ingredients. In conclusion, PFM and NFM are good sources of available protein and AAs. When fishmeal supply is limited, MBM and DDM are substitutes for fishmeal reduction in P. ussuriensis diets; at the same time, SBM and FSBM are also very acceptable protein feedstuffs. However, CSM, RSM and CGM could not be used at high levels in P. ussuriensis diets because of their inferior digestibilities.