Isolation of a Phytase-Producing Bacterial Strain from Agricultural Soil and its Characterization and Application as an Effective Eco-Friendly Phosphate Solubilizing Bioinoculant

In the present study, we isolated from agricultural soils, a Bacillus strain designated SD01N-014, expressing high-level extracellular phytase activity and investigated its potential to be used as a soil phosphate-solubilizing bioinoculant to improve soil phosphorus (P) availability and thus enhance plant growth. Soil inoculation test showed that being as an inoculant, SD01N-014 substantially enhanced soil P nutrition. Pot experiments showed that when applied to soils, SD01N-014 is capable of colonizing maize seedling roots, increasing plant-available P levels in the rhizosphere and promoting seedling growth. High tunnel trials with different vegetable plants demonstrated that SD01N-014 as a soil inoculant greatly promoted plant growth and was positively correlated with inoculum sizes. Together, these results indicate that SD01N-014 holds promising potential as a cheap, viable and eco-friendly alternative to chemical P fertilizers for increasing soil fertility and crop production in sustainable farming.     

Effects of step-up and step-down phytase regimens on performance and processing yields of male broilers from 1 to 35 d of age

An experiment was conducted to evaluate increasing or decreasing concentrations of dietary phytase on growth performance and processing yields of male broilers from 1 to 35 d of age. Treatments consisted of a positive control, a negative control (NC; less 0.14% Ca, 0.13% nonphytate P, and 0.03% Na), and 6 additional treatments based on the NC supplemented with phytase. Treatments 3 through 5 consisted of the NC diet supplemented with 500 phytase units (FTU)/kg of phytase in the starter phase that was either continued throughout the remainder of the study (treatment 3) or increased to 1,500 FTU/kg beginning in the finisher (treatment 4) or grower (treatment 5) phases. Treatment 6 had 1,500 FTU/kg of phytase throughout the study. Treatments 7 and 8 had 1,500 FTU/kg in the starter and decreased to 500 FTU/kg in the finisher or grower phases, respectively. At 35 d of age, broilers fed diets containing 1,500 FTU/kg of phytase had increased BW gain compared with birds fed diets formulated to contain 500 FTU/kg of phytase. Increasing phytase concentration between the starter and grower phases or decreasing phytase concentration between the grower and finisher phases negatively affected FCR from 1 to 35 d of age. Phytase supplementation did not affect weight and yield of carcass characteristics. Therefore, dietary phytase concentration should not be varied throughout production for optimum growth performance.     

玉米-杂粕型日粮中添加不同植酸酶对蛋鸡生产性能和蛋品质及钙磷代谢的影响

选取720只24周龄海兰褐蛋鸡,随机分为8个处理,每个处理6个重复,每个重复15只鸡。8个处理分别为正对照组(PC)饲喂有效磷为0.36%的正常基础日粮,负对照组(NC)饲喂有效磷为0.16%的低磷日粮,其余试验组是在NC组日粮基础上分别添加两种植酸酶A或B各125、250、500 U/kg。其中植酸酶A是从大肠杆菌中提取,植酸酶B是从里氏木霉中提取。试验期为12周。结果表明:与PC组比较,NC和B500组产蛋率、平均蛋重和日采食量显著降低(P0.05),其他各添加植酸酶组恢复到PC组水平,各添加植酸酶组料蛋比和蛋品质无显著变化(P0.05);与PC组相比,NC组胫骨灰分、胫骨磷和血浆磷含量显著降低(P0.05),添加2种植酸酶组(除B500组外)恢复到PC组水平,B500组血浆磷含量显著低于PC组(P0.05),各试验组胫骨钙含量和血浆钙含量与PC组无显著差异(P0.05)。结果提示,低磷玉米-杂粕型日粮中添加2种植酸酶都能够缓解低磷引起的负效应,但2种植酸酶的最佳添加量不同,本试验条件下日粮中添加125 U/kg的植酸酶B或250 U/kg的植酸酶A都能满足蛋鸡对无机磷的需求。     

不同磷水平和钙磷比饲粮添加植酸酶对妊娠母猪养分表观消化率的影响

本试验旨在研究不同磷水平和钙磷比饲粮添加植酸酶对妊娠母猪养分消化率的影响。试验选择遗传背景一致、2~3胎次(预产期前28 d)、体重为(221.91±7.41)kg、预产期相近的健康"长×大"二元杂交母猪36头,按照胎次、预产期均衡原则随机分为9个组,每个组4个重复。对照组饲喂基础饲粮(钙、磷水平分别为7.26、5.81 g/kg);组2为基础饲粮添加植酸酶;组3~5保持钙水平(7.26 g/kg)不变添加植酸酶,减少无机磷添加至对照组的75%、50%、25%[钙磷比为(1.35~1.63)∶1.00];组6~9保持钙磷比不变(1.25∶1.00)添加植酸酶,减少无机磷添加至对照组的75%、50%、25%、0。结果表明:1)正常磷水平下,添加植酸酶显著提高了干物质、粗灰分、有机物和磷的表观消化率(P0.05),极显著降低了粪中磷的排泄量(P0.01),有降低铜、锌排泄量的趋势(P0.10)。与对照相比,低磷水平添加植酸酶极显著提高了干物质、粗灰分、有机物、磷、钙、铁、铜、锌的表观消化率(P0.01),显著降低了粪中磷、钙、铁、铜、锌的排泄量(P0.05)。2)饲粮在保持钙水平不变或钙磷比不变的条件下添加植酸酶,随磷添加量的降低,干物质、粗灰分、有机物、磷、铁、铜、锌的表观消化率均呈线性或二次升高(P0.05),粪中磷、铁、铜、锌的排泄量呈线性或二次降低(P0.01),保持钙磷比不变,钙、粗蛋白质的表观消化率显著提高(P0.05)。3)以表观可消化磷为标准,饲粮保持钙水平不变和钙磷比不变添加植酸酶可分别替代0.91、1.27 g/kg的无机磷,减少24.70%、34.48%的粪磷排泄量。4)钙磷比不变(1.25∶1.00)组饲粮的养分表观消化率极显著高于钙水平不变[高钙磷比(1.35~1.63)∶1.00]组饲粮(P0.01),且粪矿物质的排泄量极显著减少(P0.01)。综上所述:1)在本研究条件下,在低无机磷、低钙磷比(1.25∶1.00)饲粮中,添加植酸酶提高养分消化率的效果更显著。2)以表观可消化磷为标准,母猪饲粮的钙磷比为1.25∶1.00时,添加植酸酶可减少1.27 g/kg的磷添加。     

Phosphorus utilization in juvenile Clarias gariepinus fed phytase-supplemented diets based on soya bean (oil-extracted) and full-fat (roasted): A comparison

The effect of microbial phytase on phosphorus utilization in juvenile Clarias gariepinus (initial fish body weight 11.55 ± 0.2 g) was tested on two different diets based on oil-extracted soya bean (Experiment 1) and roasted soya bean meal (Experiment 2) using a 5 × 5 experimental design for 84 days. The basal isonitrogenous and isocaloric diets for oil-extracted and roasted soya bean were formulated to replace fish meal at 25% (S1E), 50% (S2E), 75% (S3E), 100% (S4E); and 25% (S1), 50% (S2), 75% (S3), 100% (S4), respectively. Each treatment was replicated four times. Microbial phytase was supplemented in each replicate at 250 FTU/g (P1), 500 FTU/g (P2), 750 FTU/g (P3), and 1,000 FTU/g (P4). Basal controls, which included a fish meal-based diet (S0), were not supplemented with phytase (P0). The result in Experiment 1 showed that there was a significant increase in whole-body protein and reduction in fat with phytase compared to a diet without phytase (P < 0.05). Serum total protein declined significantly with phytase supplementation (P < 0.05). Serum phosphorus and glucose were higher with phytase supplementation compared to control (P < 0.05). Bone minerals declined significantly with increasing level of soya bean compared to fish meal diet (P < 0.05). In Experiment 2, serum phosphorus was improved with phytase compared to control with no phytase (P > 0.05). A significant reduction in whole-body protein and increase in fat was observed for fish fed phytase diets compared to diets with no phytase, regardless of soya bean level (P < 0.05); however, ash content was improved with phytase (250 FTU/g) compared to control (P < 0.05). Phytase supplementation improved bone phosphorus (250 FTU/g), calcium (250 FTU/g), magnesium (250–500 FTU/g), and zinc (250–1,000 FTU/g) compared to control (P < 0.05). In conclusion, the research has demonstrated that improved bone phosphorus (P) and growth could be achieved with the supplementation of dietary phytase.     

Study on the Production of Porcine Phytase Transgenic Embryos with Somatic Cell Nuclear Transfer

In order to get porcine phytase salivary glands specific express cloned embryos,this study used phytase specific expressed plasmid DNA (including the promotor and terminater sequences of parotid secretary protein (PSP) gene,Neo selected gene and EGFP report gene and high enzymatic activity phytase gene (appA)) and transfected it into porcine fetal fibroblast cells with liposome.The stably transfected cell lines were screened by G418 selection,and positive phytase transgenic embryos were constructured by somatic cell nuclear transfer.The results showed that the phytase plasmid of this experiment could be used to carry on the cell selection,and the more smaller of plasmid DNA size,the more higher transfected efficiency was got.The plasmid YM6552 with 14.89 kb size only got 7.1% transfected efficiency,while EGPF plasmid got 43.4%.In the formation of single cell colony,pYN3600 (more smaller DNA size) got 25 colony numbers,and 14 of them expressed green fluorescent protein,11 of them were phytase PCR positive.They were much higher than those of YM6552 (19,8 and 6).All the reconstructed transgenic embryoes from the positive cell lines were expressed green fluorescent protein.Though the in vitro development ability of transgenic cloned embryos were a little lower than that of control group,there were no significant differences between them (P>0.05).In conclusion,all the phytase plasmid,cell selection method and nuclear transfer techniques could be used to produce porcine phytase salivary glands specific express embryos.     

Soy protein concentrate as a substitute for fish meal in diets for juvenile Acanthopagrus schlegelii: effects on growth,phosphorus discharge and digestive enzyme activity

The current high demand and cost of fish meal (FM) necessities the evaluation of alternative plant protein ingredients in diets of farmed marine fish. A 56‐day feeding trial was performed to study the effects of replacement of FM with soy protein concentrate (SPC) in diets of Acanthopagrus schlegelii. Diets were prepared at levels of 0%, 30%, 47.5%, 65%, 82.5% and 100% SPC, respectively, replacing FM. The results indicated no significant differences (p > .05) in % weight gain (WG) and specific growth rate of fish fed S30% to S82.5% diets compared with the control diet while, further inclusion at 100% significantly depressed growth performance. SPC inclusion and phytase supplementation significantly affected the phosphorous discharge (P‐load) showing lowest value (3.83 ± 0.53 g/kg WG) in S100% compared to control (14.79 ± 0.37 g/kg WG) and in fish fed S30% diet (13.24 ± 0.89 g/kg WG) (p < .05). The results of this study showed that FM could be substituted up to 82.5% by SPC in the diet of Acanthopagrus schlegelii fingerlings (5.53 ± 0.12 g) without any adverse effects. Phytase supplementation SPC based diets could be effective in reducing the phosphorus load in the aquatic culture environment.