Implication and challenges of direct-fed microbial supplementation to improve ruminant production and health

Direct-fed microbials(DFMs)are feed additives containing live naturally existing microbes that can benefit animals’health and production performance.Due to the banned or strictly limited prophylactic and growth promoting usage of antibiotics,DFMs have been considered as one of antimicrobial alternatives in livestock industry.Microorganisms used as DFMs for ruminants usually consist of bacteria including lactic acid producing bacteria,lactic acid utilizing bacteria and other bacterial groups,and fungi containing Saccharomyces and Aspergillus.To date,the available DFMs for ruminants have been largely based on their effects on improving the feed efficiency and ruminant productivity through enhancing the rumen function such as stabilizing ruminal pH,promoting ruminal fermentation and feed digestion.Recent research has shown emerging evidence that the DFMs may improve performance and health in young ruminants,however,these positive outcomes were not consistent among studies and the modes of action have not been clearly defined.This review summarizes the DFM studies conducted in ruminants in the last decade,aiming to provide the new knowledge on DFM supplementation strategies for various ruminant production stages,and to identify what are the potential barriers and challenges for current ruminant industry to adopt the DFMs.Overall literature research indicates that DFMs have the potential to mitigate ruminal acidosis,improve immune response and gut health,increase productivity(growth and milk production),and reduce methane emissions or fecal shedding of pathogens.More research is needed to explore the mode of action of specific DFMs in the gut of ruminants,and the optimal supplementation strategies to promote the development and efficiency of DFM products for ruminants.

Isolation, screening and identifi cation of Bacillus spp. as direct-fed microbial candidates for aflatoxin B_1 biodegradation

Objective: To evaluate the ability of Bacillus spp. as direct-fed microbials(DFM) to biodegrade al atoxin B1(AFB1) by using an in vitro digestive model simulating in vivo conditions.Methods: Sixty-nine Bacillus isolates were obtained from intestines, and soil samples were screened by using a selective media method against 0.25 and 1.00 μg/m L of AFB1 in modii ed Czapek-Dox medium. Plates were incubated at 37 °C and observed every two days for two weeks. Physiological properties of the three Bacillus spp. candidates were characterized biochemically and by 16 S r RNA sequence analyzes for identii cation. Tolerance to acidic p H, osmotic concentrations of Na Cl, bile salts were tested, and antimicrobial sensitivity proi les were also determined. Bacillus candidates were individually sporulated by using a solid fermentation method and combined. Spores were incorporated into 1 of 3 experimental feed groups: 1) Negative control group, with unmedicated starter broiler feed without AFB1; 2) Positive control group, with negative control feed contaminated with 0.01% AFB1; 3) DFM treated group, with positive control feed supplemented with 109 spores/g. After digestion time(3:15 h), supernatants and digesta were collected for high-performance liquid chromatography l uorescence detection analysis by triplicate.Results: Three out of those sixty-nine DFM candidates showed ability to biodegrade AFB1 in vitro based on growth as well as reduction of l uorescence and area of clearance around each colony in modii ed Czapek-Dox medium which was clearly visible under day light after 48 h of evaluation. Analysis of 16S-DNA identii ed the strains as Bacillus amyloliquefaciens, Bacillus megaterium and Bacillus subtilis. The three Bacillus strains were tolerant to acidic conditions(p H 2.0), tolerant to a high osmotic pressure(Na Cl at 6.5%), and were able to tolerate 0.037% bile salts after 24 h of incubation. No signii cant dif erences(P > 0.05) were observed in the concentrations of AFB1 in neither the supernatants nor digesta samples evaluated by highperformance liquid chromatography with l uorescence detection between positive control or DFM treated groups. Conclusions: In vitro digestion time was not enough to confirm biodegradation of AFB1. Further studies to evaluate the possible biodegradation ef ects of the BacillusDFM when continuously administered in experimentally contaminated feed with AFB1, are in progress.

Direct-fed microbes: A tool for improving the utilization of low quality roughages in ruminants

For many years, ruminant nutritionists and microbiologists have been interested in manipulating the microbial ecosystem of the rumen to improve production efficiency of different ruminant species. Removal and restriction of antibiotics subtherapeutic uses from ruminant diets has amplified interest in improving nutrient utilization and animal performance and search for more safe alternatives. Some bacterial and fungal microorganisms as a direct-fed microbial（DFM） can be the most suitable solutions. Microorganisms that are commonly used in DFM for ruminants may be classified mainly as lactic acid producing bacteria（LAB）, lactic acid utilizing bacteria（LUB）, or other microorganism＇s species like Lactobacillus, Bifidobacterium, Enterococcus, Streptococcus, Bacillus, Propionibacterium, Megasphaera elsdenii and Prevotellabryantii, in addition to some fungal species of yeast such as Saccharomyces and Aspergillus. A definitive mode of action for bacterial or fungal DFM has not been established; although a variety of mechanisms have been suggested. Bacterial DFM potentially moderate rumen conditions, and improve weight gain and feed efficiency. Fungal DFM may reduce harmful oxygen from the rumen, prevent excess lactate production, increase feed digestibility, and alter rumen fermentation patterns. DFM may also compete with and inhibit the growth of pathogens, immune system modulation, and modulate microbial balance in the gastrointestinal tract. Improved dry matter intake, milk yield, fat corrected milk yield and milk fat content were obtained with DFM administration. However, the response to DFM is not constant; depending on dosages, feeding times and frequencies, and strains of DFM. Nonetheless, recent studies have supported the positive effects of DFM on ruminant performance.

Bacterial direct-fed microbials fail to reduce methane emissions in primiparous lactating dairy cows

Direct-fed microbials(DFM) are considered as a promising technique to improve animal productivity without affecting animal health or harming the environment.The potential of three bacterial DFM to reduce methane(CH4)emissions,modulate ruminal fermentation,milk production and composition of primiparous dairy cows was examined in this study.As previous reports have shown that DFM respond differently to different diets,two contrasting diets were used in this study.Eight lactating primiparous cows were randomly divided into two groups that were fed a corn silage-based,high-starch diet(HSD) or a grass silage-based,high-fiber diet(HFD).Cows in each dietary group were randomly assigned to four treatments in a 4 × 4 Latin square design.The bacterial DFM used were selected for their proven CH4-reducing effect in vitro.Treatments included control(without DFM) and 3 DFM treatments: Propionibacterium freudenreichii 53-W(2.9 × 10^10 colony forming units(CFU)/cow per day),Lactobacillus pentosus D31(3.6 × 10^11 CFU/cow per day) and Lactobacillus bulgaricus D1(4.6 × 10^10 CFU/cow per day).Each experimental period included 4 weeks of treatment and 1 week of wash-out,with measures performed in the fourth week of the treatment period.Enteric CH4 emissions were measured during 3 consecutive days using respiration chambers.Rumen samples were collected for ruminal fermentation parameters and quantitative microbial analyses.Milk samples were collected for composition analysis.Body weight of cows were recorded at the end of each treatment period.Irrespective of diet,no mitigating effect of DFM was observed on CH4 emissions in dairy cows.In contrast,Propionibacterium increased CH4 intensity by 27%(g CH4/kg milk) in cows fed HSD.There was no effect of DFM on other fermentation parameters and on bacterial,archaeal and protozoal numbers.Similarly,the effect of DFM on milk fatty acid composition was negligible.Propionibacterium and L.pentosus DFM tended to increase body weight gain with HSD.We conclude that,contrary to the effect previously observed in vitro,bacterial DFM Propionibacterium freudenreichii 53-W,Lactobacillus pentosus D31 and Lactobacillus bulgaricus D1 did not alter ruminal fermentation and failed to reduce CH4 emissions in lactating primiparous cows fed high-starch or high-fiber diets.

Effect of Yeast Culture and Direct-Fed Microbes on the Growth Performance and Rumen Fermentation of Weaner Lambs

The effects of yeast culture and directfed microbes on the growth performance of weaner lambs was examined. Thirty-two Hu lambs with inihtial weight of 22.20 （ ±0.75 ） kg were randomly assigned to one of four treatments： basal diet without additive （control）, added with yeast culture at 15 g/head/d （YEC）, YEC plus Bacillus licheniformis preparation at 2. 3 g/head/d （YBL） or plus Clostridium butyricum preparation at 2. 3 g/head/d （YCB）. The feeding trial lasted 75 d with 15 d for adaptation. Feed intake was not influenced （P 〉0. 05） by treatment. Average daily gain of growing lambs was 102, 114, 90, and 89 g/d in control, YEC, YBL, and YCB, respectively, with no significant difference （P 〉 0.05） among treatments, but the carcass weight of YEC lambs was significantly higher （P 〈 0.05） than that of other treatments. Total volatile fatty acids and acetate to propionate ratio in the rumen were unaffected, although the butyric acid concentration was higher （ P 〈 0.05 ） in the ru men fluid of YCB lambs compared with YEC lambs and slightly higher （ P 〉 0. 05 ） than in controls and YBL lambs. Solid-associated fungi population relative to total rumen bacteria 16S ribosomal DNA was significantly lower （ P 〈 0. 05 ） in YBL lambs （3.55） compared with those on YCB （23.12）. There was little difference in blood glucose and plasma urea-N concentrations among the treatments. Blood concentrations of creatinine and globulin were significantly higher （P 〈0.05） in YBL lambs, compared with the control and YEC-fed animals, and no difference with YCB lambs. Total protein and triglycerides in blood were significantly （P 〈 0.05） higher in YBL lambs, compared with controls. These serum biochemical parameters suggest that treatment increased amounts of absorbable protein but not efficiency of protein utilization and in YBL and YCB lambs. The results indicated that yeast culture improve growth performance, while little advantage could be expected from combining yeast culture with either the B. licheniformis preparation or C. butyricum preparation. More research using lower doses of B. licheniformis prepara- tion or C. butyricum preparation in combination with yeast culture is warranted.

Prospective use of bacteriocinogenic Pediococcus pentosaceus as direct-fed microbial having methane reducing potential

Direct-fed microbials（DFM）, generally regarded as safe status, are successfully used in improving rumen ecology, gastro-intestinal health, feed efficiency, milk production and growth rate in ruminants. On the other hand, methanogenesis in rumen, which accounts for a significant loss of ruminant energy and increased greenhouse gas in environment, is of great concern, therefore, use of DFM for improving productivity without compromising the animal health and ecological sustainability is encouraged. The present study was conducted to investigate the methane reducing potential of bacteriocinogenic strain Pediococcus pentosaceus-34. Since, the culture showed no hemolysis on blood agar and DNase activity, hence, it was considered to be avirulent in nature, a prerequisite for any DFM. The culture also showed tolerance to pH 5.0 for 24 h with 0.5% organic acid mixture, whereas when given a shock for 2 h at different p H and organic acids concentrations, it showed growth at pH 3.0 and 4.0 with 0.1 and 1.0% organic acids, respectively, as having good animal probiotics attributes. The total gas production was significantly（P〈0.05） higher in live pedicoccal culture（LPC） and dead pedicoccal culture（DPC） both with wheat straw, when compared to the control. In sugarcane bagasse, gas production was significantly lower（P〈0.05） with LPC compared to the control and DPC both. Methane was reduced by the inclusion of LPC in sugarcane bagasse（0.07 mL CH4 mg–1 dry matter digestibility） with no effect on other rumen fermentation parameters. However, with wheat straw and LPC total gas, in vitro dry matter digestibility, total volatile fatty acids increased significantly but no reduction in methane production was observed in comparison to the control. Therefore, further research is warranted in this direction, if the bacteriocinogenic strains can be used as DFM for ruminants to improve the ruminant productivity.

Effect of Direct-Fed Microbial Supplementation on Pathogenic <i>Escherichia coli</i>Fecal Shedding, Live Performance, and Carcass Characteristics in Feedlot Steers

Three experiments were conducted to evaluate direct-fed microbial (DFM) supplementation on live performance, carcass characteristics, and fecal shedding of E. coli in feedlot steers. In Exp. 1, 400 steers (BW = 348 kg) were assigned to treatments: CON = lactose carrier only, BOV = P. freudenreichii (NP24) + L. acidophilus (NP51), BOVD = P. freudenreichii (NP24) + L. acidophilus (NP51), and COMB = BOV fed for the first 101 d on feed, followed by BOVD for the final 28 d prior to harvest. In Exp. 2 (n = 1800;BW = 354 kg) and Exp. 3 (n = 112;BW = 397 kg), steers were utilized in a randomized complete block design and assigned to DFM treatments using low dose and high dose, respectively. Fecal samples were collected prior to harvest and analyzed for E. coli serogroups. In Exp. 1, DFM reduced (P < 0.01) the concentration of E. coli O157. Prevalence of O157 was reduced by BOVD supplementation in Exp. 2 and 3 (P < 0.01 and P = 0.08, respectively), and concentration of E. coli O157 in positive samples was reduced in both experiments where enumeration was performed (P ≤ 0.02). Weighted mean differences across the three experiments were equal to a 33% reduction in the prevalence of E. coli O157:H7 in BOVD treated cattle. A significant reduction in prevalence of O26, O45, O103, and O121 was observed in Exp. 2 (P ≤ 0.03). These results indicate that high levels of L. acidophilus (NP51) may represent an effective pre-harvest food safety intervention to reduce fecal shedding of several E. coli serogroups.

瘤胃源粪臭素降解菌的分离鉴定及其降解特性研究

【目的】旨在从绵羊瘤胃中分离粪臭素降解菌,并评估其粪臭素降解能力和生长性能,以期开发适用于反刍动物降臭的直接饲喂微生物。【方法】以绵羊瘤胃液为分离来源,使用含有粪臭素的MSM培养基进行富集和分离,通过细菌菌落形态观察进行初步分类;应用16S rRNA基因扩增、测序及系统发育分析进行物种鉴定;绘制菌株生长曲线,通过HPLC技术测定粪臭素降解曲线。【结果】从绵羊瘤胃液中分离出25株粪臭素降解菌,经菌落形态鉴定选出11株代表菌株进行后续研究。物种鉴定结果显示,MSML2和MSML6属于枯草芽孢杆菌(Bacillus subtilis),MSML4、MSML5、MSML7和MSML10属于阿氏普里斯特氏菌(Priestia aryabhattai),MSML3和MSML11属于污染伯克霍尔德氏菌(Burkholderia contaminans),MSML1、MSML8和MSML9属于成都假单胞菌(Pseudomonas chengduensis)。其中,MSML5生长速度最快,在约12 h后进入稳定期,稳定期菌体浓度最高;MSML3和MSML8在前16 h生长缓慢,32 h后进入稳定期。在粪臭素降解方面,MSML2粪臭素降解效率最高,48 h内的降解率达到23.03%,其次是MSML7、MSML8和MSML10,降解率均超过20%。【结论】成功从绵羊瘤胃中分离获得25株粪臭素降解菌,涉及4个物种,首次报道了具备粪臭素降解能力的枯草芽孢杆菌、阿氏普里斯特氏菌和成都假单胞菌,为直接饲喂反刍动物的菌剂开发提供了宝贵的菌株资源。

雨养农业区黄芩轻简化膜间直播种植技术规程

为进一步推进甘肃省雨养农业区黄芩种植机械化、精量化和轻简化生产,结合多年的试验示范和生产实践,从范围、规范性引用文件、术语和定义、生产环境、栽培技术、田间管理、采收与产地加工等方面总结出了雨养农业区黄芩轻简化膜间直播种植技术规程。

双定子无刷双馈风力发电机无速度传感器反步法直接功率控制

针对双定子无刷双馈发电机,将内外功率绕组及内外控制绕组分别同相串联以保证电磁一致性。基于Lyapunov和Popov稳定性理论,采用反步法和模型参考自适应系统设计一种无速度传感器的反步法直接功率控制策略,利用空间矢量脉宽调制(SVPMW)以固定开关频率驱动控制绕组机侧变流器IGBT,以减小功率绕组功率波动及电流总谐波畸变率。通过12/8极50 kW样机控制系统的特性仿真研究,结果表明:所提控制策略在亚同步~超同步的速度变化范围内,可实现变速恒频发电、最大功率跟踪、无功功率及单位功率因数控制,电流总谐波畸变率约为2%。

基于直接磁链跟踪控制的无刷双馈电机无Crowbar对称低电压穿越方法

低电压穿越(low-voltage ride through,LVRT)能力是并网型风电机组重要的性能指标之一。该文提出一种基于直接磁链跟踪控制的无刷双馈感应发电机(brushlessdoubly fed induction generator,BDFIG)无Crowbar对称低电压穿越方法。该方法基于开关表,通过直接选定电压空间矢量来控制无刷双馈电机控制绕组(control winding,CW)磁链的幅值和相位,以迅速抑制低电压穿越期间控制绕组的感应过电压和由此导致的过电流,同时快速向电网注入动态无功电流以支撑电压恢复。该方法无需坐标变换、系统结构简单、磁链动态响应迅速。样机实验结果表明,低电压穿越过程中,有功恢复及无功功率动态响应满足国标相关要求,且控制绕组电流得到有效抑制,验证了所提控制策略的有效性。

公转和自转协同下绕包机转速自动控制研究

绕包机公转和自转过程中可能会产生非线性影响,为了实现转速有效控制,该文提出一种公转和自转协同下绕包机转速自动控制方法。构建神经网络控制模型,将公转速度和自转速度作为输入,通过输出层输出控制规律;利用反向传播算法,通过调节网络权值和引入动量权值对神经网络模型进行改进,以此提升公转和自转协同控制效果;利用双馈定向矢量方法实施绕包机转速自动控制,在满足绕包机定子和转子对应电压矢量和磁链矢量条件的基础上,完成最终的转速自动控制。在不同实验条件下,所提方法对输入信号的实际跟踪效果较好,且不易受影响因素干扰。

基于LESO的DFIG-GSC反馈线性化直接功率控制

为了提高双馈风力发电机网侧变换器(DFIG-GSC)这个强耦合非线性系统的性能,提出了一种反馈线性化直接功率控制策略。首先在αβ坐标系下建立以有功功率、无功功率为变量的DFIG-GSC的数学模型,针对模型中的强耦合项采用反馈线性化进行解耦,得到其线性模型,再结合滑模控制理论设计出功率内环的控制律。由于功率内环控制律中未建模部分和不确定因素会对控制效果产生影响,采用线性扩张状态观测器对其进行观测并对其控制量进行修正。同时电压外环采用滑模控制来提高直流侧电压的响应速度。最后搭建仿真模型与传统PI控制以及直接功率控制进行比较,验证了所提控制策略的合理性和有效性。

开绕组无刷双馈风力发电机直接功率控制容错策略

针对无刷双馈发电机提出了一种控制绕组采用开绕组拓扑(OW-BDFG)的馈电方案,将三相控制绕组两端打开分别连接变流器构成双两电平变流器(D-TLCs)馈电拓扑,对机侧变流器功率器件IGBT因故障切除后的容错控制策略进行深入分析,提出了功率偏差比较直接功率控制(PEC-DPC)的D-TLCs电压空间矢量优化方案。对比分析了8/4极25kW样机的仿真及实验结果,表明:当机侧变流器1个或2个IGBT被切除后,所提OW-BDFG的PEC-DPC仍可实现变速恒频、最大功率点跟踪及单位功率因数控制,进一步提高了该发电机系统的可靠性和鲁棒性。

双馈感应电机无速度传感器直接转矩控制方法

风电机组双馈感应发电机运行中,由于参数变化的不确定性导致其无速度传感器直接转矩控制输出值出现异常波动,为保证双馈感应电机的频率稳定性,提出一种双馈感应电机无速度传感器直接转矩控制方法。计算转子磁链同步转速和转差速度,得到发电机的转速观测值;分析转子绕组电压空间矢量,建立磁链模型并构建电磁矩阵,在矩阵中计算定子绕组和转子绕组的磁势矢量,通过二者的矢量积得到电机的电磁转矩;转速和转矩共同组成直接转矩控制系统,实现对电机的速度调节和控制。仿真分析结果表明:采用所提方法时,在双馈感应电机启动后转速迅速增加,且在1.2 s后转速始终保持为1400 r/min,可保证转子磁链的运行轨迹接近于圆形,直接转矩控制效果平稳。

基于阻抗幅值波动差异的新型参数识别方向元件

针对故障分量方向元件在大规模双馈风机并网不适用的问题,提出了基于正序阻抗幅值波动差异的新型参数识别方向元件。首先,将风电场正序阻抗波动量化,分析阻抗波动对正序故障分量方向元件影响,得到正序故障分量方向元件在风电场送出线路的拒动边界。其次,基于Prony算法结合带通滤波器提取工频量,分析送出线路风电侧正、反方向故障正序阻抗幅值变化特征。最后,引入去趋势波动分析(detrended fluctuation analysis, DFA)计算波动函数,构造新型参数识别方向元件动作判据,实现故障方向判别。仿真结果表明,提出的参数识别方向元件不受风电场运行工况、故障类型和系统振荡的影响。当过渡电阻达到100Ω时,仍能正确判别故障方向,具有较强的抗干扰性能,适用于含风电接入的弱馈型电力系统。

基于EMPC的无刷双馈发电机直接功率控制

针对无刷双馈发电机模型复杂、耦合性强的问题,提出一种基于扩张状态观测器显式模型预测的直接功率控制策略。通过研究定子控制绕组侧电压与功率绕组侧有功功率和无功功率的关系,推导基于d-q旋转坐标系的直接功率控制状态方程,对功率实现解耦控制。针对模型预测控制在线计算的局限性和系统中存在的扰动因素,提出一种基于扩张状态观测器的显式模型预测控制。该控制策略通过扩张状态观测器对扰动项进行补偿,结合显式模型预测控制将模型预测控制中的优化问题移到“线下解决”,该控制策略提高了在线计算速度和精度,同时扩大了模型预测控制的应用范围。详细介绍了该控制策略的设计原理,并对30 kW低功率无刷双馈风力发电机的控制策略进行了仿真验证。结果表明,该控制方案性能良好,能够提高无刷双馈电机的抗扰动能力,特别是提高了转速和功率变化时的稳定性和鲁棒性。

壳聚糖饲用微生物制剂对湘黄鸡免疫指标和肠道菌群的影响

在不用药物、不接种疫苗的条件下,于饮水中添加0(CK),0.5%,1.0%和1.5%的以壳聚糖为培养底物研制的壳聚糖饲用微生物制剂饲喂湘黄鸡,探讨其对湘黄鸡免疫指标和肠道菌群的影响效果.结果表明,试验组脾脏指数有上升趋势(P<0.25),法氏囊指数和胸腺指数显著提高(P<0.05);试验组血液中IgG含量呈上升趋势(P<0.25),T3,T4和IgA含量也显著提高(P<0.05或P<0.01);试验组盲肠中大肠杆菌显著减少,而乳酸杆菌显著增加(P<0.01).同时在日增重和育成率上,试验组提高率达9.31%(P<0.01)和7.01%(P<0.05),表明该制剂通过促进免疫器官发育,增强T3,T4,IgA,IgG分泌和改善肠道微生态环境,从而提高试鸡的抗病力和生长性能.

利用PCR-DGGE技术快速检测鉴定饲用微生物

针对目前国内外对饲用微生物添加剂中多种微生物的检测缺乏简捷有效手段这一现实状况,通过PCR-DGGE方法建立了地衣芽孢杆菌Bacillus licheniformis、粪肠球菌Enterococcus faecalis、植物乳杆菌Lactobacillus planta-rum、干酪乳杆菌Lactobacillus casei4种标准菌株的分子指纹图谱,建立了一套这4种微生物的快速同步检测技术,并用该检测技术对市场上一种饲用微生物添加剂进行分析,结果判定其含有6种优势菌群,包括植物乳杆菌和干酪乳杆菌.本技术还可以推广应用到目前国家允许添加的其他16种饲用微生物的检测,为饲用微生物添加剂产品质量监控提供了技术支撑.

饲用微生物评价研究进展

随着饲用微生物产品的发展,饲用微生物评价方法相应而生。通过综述饲用微生物品种鉴定,耐酸性和耐胆盐一般特性评价,生产功效评价,粘附作用、抑菌作用和免疫刺激作用评价的研究进展,期望为建立完善的饲用微生物评价体系提供参考。