益生菌安全性和有效性评价方法的研究进展

益生菌是指一类活的、摄入足够量就能够对人体产生有益作用的微生物,目前广泛应用于食品和饲料工业领域。随着市场上商品化益生菌的不断出现,如何评价一个菌株的安全性及有效性成为人们关注的问题。目前,益生菌的评价主要从安全性和有效性入手,安全性和有效性评价又包括体内和体外的评价。很多企业在开发益生菌产品时很少对菌株做出全面的安全性评价,而且各国研究者采用的评价指标和评价方法也不尽相同。对于有效性评价,同样也没有统一的评价标准。针对目前益生菌评价存在的问题,亟需建立一套完善的评价方法,对益生菌的安全性和有效性进行系统、科学地评价。作者从益生菌可能产生的潜在危害及功效方面出发,着重论述了益生菌的溶血性、毒力因子、抗生素抗性和产有毒代谢产物的体外安全性评价方法,耐酸、耐胆盐、耐人工胃肠液、胃肠黏膜黏附性和抑菌活性的体外有效性评价方法,及体内的安全性和有效性评价方法的研究进展,期望为建立完善的益生菌评价技术体系和标准提供参考。     

广西猪源益生菌的分离鉴定与生物学特性研究

为了解广西地区猪粪便与微生态发酵饲料中益生菌的存在情况及合理开发和利用猪源益生菌,本试验共采集了15份猪粪便样品和5份益生菌发酵饲料并对其进行益生菌的分离,研究益生菌的形态学特性、生理生化特性及16S rRNA分子序列,进行生长曲线、耐胆盐试验、温度敏感试验、模拟胃肠道耐受试验、耐药性试验、体外抑菌试验、小鼠安全性试验、体内/外抑菌试验,对分离菌株进行生物学特性的研究分析。结果显示,本试验共分离鉴定出6种、29株益生菌,分别为1株屎肠球菌（C2）、2株乳酸肠球菌（C1、C3）、3株植物乳杆菌（R20、R30、R37）、4株干酪乳杆菌（R41~R44）、7株枯草芽孢杆菌（K1~K7）和12株蜡样芽孢杆菌（Y1~Y12）。从中筛选出6株具有生长性能好、耐胆盐、耐高温、对胃肠道耐受、体外抑菌能力强的益生菌：乳酸肠球菌C1、屎肠球菌C2、植物乳杆菌R20、干酪乳杆菌R41、枯草芽孢杆菌K1和蜡样芽孢杆菌Y3。将筛选出的具有优良特性的菌株进行小鼠安全性试验,结果表明,在10⁹ CFU/mL浓度下灌胃6株益生菌对小鼠是安全无毒害的。与此同时,C2、R20、R41、Y3能极显著提高小鼠日增重（P<0.01）,K1能显著提高小鼠日增重（P<0.05）。体内抑菌试验结果表明,C2、R20、K1能降低沙门氏菌G21感染的小鼠的死亡率,有一定程度的体内抑菌作用。各项试验结果表明,R20和K1可作为猪源益生菌的选择菌株。     

Study and use of the probiotic Lactobacillus reuteri in pigs:a review

Probiotics are living microorganisms that provide a wide variety of health benefits to the host when ingested in adequate amounts.The bacterial strains most frequently used as probiotic agents are lactic acid bacteria,such as Lactobacillus reuteri,which is one of the few endogenous Lactobacillus species found in the gastrointestinal tract of vertebrates,including humans,rats,pigs and chickens.L reuteri is one of the most well documented probiotic species and has been widely utilized as a probiotic in humans and animals for many years.Initially,L reuteri was used in humans to reduce the incidence and the severity of diarrhea,prevent colic and necrotic enterocolitis,and maintain a functional mucosal barrier.As interest in alternatives to in-feed antibiotics has grown in recent years,some evidence has emerged that probiotics may promote growth,improve the efficiency of feed utilization,prevent diarrhea,and regulate the immune system in pigs.In this review,the characteristics of L reuteri are described,in order to update the evidence on the efficacy of using L.reuteri in pigs.     

Probiotics--possibilities and limitations of their application in food, animal feed, and in pharmaceutical preparations for men and animals]

Probiotics are cultures of special microorganisms, which have been used as feed additives since the seventies of the past century and already since the twenties in food specimen and in pharmaceuticals. The definition of "Probioticum" was formulated in 1974 simultaneously with the use of living cultures in feed for various animals in order to substitute the application of nutritive antibiotics or chemotherapeutics. In the meantime probiotics are applied not only as feed supplements or pharmaceuticals but increasingly in suitable food specimens such as dairy products, fruit juices, chocolates, and even meat products. Of course, heating of such products prior to consumption or application must be ruled out. The selection of a suitable strain of a microorganism can be regarded as the primary requirement for the use as a probiotic. These cultures must be able to pass the stomach-duodenum barrier in a viable state and to multiply at the site of destination in the intestine. Additionally, they must be capable of producing antagonistic metabolites against a dominating saprophytic microflora resulting in a competitive growth. These abilities are common among lactic acid bacteria, e.g. lactobacilli and bifidobacteria. Yet, intended autochthonous species do not possess very good technological features for the enrichment in food specimens, because their viability will decrease rapidly under unfavourable conditions as it is the case with mineral supplements in feed or with low acidity in fermented dairy products. Therefore, some other microorganisms like spore-forming species or yeast cultures were introduced as probiotic components. These possess some similar features which render them suitable for probiotic use. But their physiological and ecological traits do not qualify them as probiotics of first choice. The special efficacy of probiotics must be strictly verified in animal nutrition due to restrictive EC-regulations, in pharmacy due to legal restraints, and in food applications in accordance with food law regulations. Safety aspects are considered very restrictively in feed applications, in the food and pharmaceutical sector they should be in accordance with the intended purpose of "fulfilling health claims". In the presented review the different requirements for the application of probiotics in animal nutrition, in food, and in pharmaceuticals will be provided. The special effect of competitive exclusion of pathogenetic and toxinogenic microorganisms in fowl performance are mentioned and even the application of probiotics as marine aquacultures of fish and Crustaceae will be included. Furthermore, the safety aspects resulting from the tremendous amount of industrially produced cultures which are distributed into the environment will be discussed. In conclusion it may be pointed out that probiotics may serve to partially replace the presently reduced or even prohibited application of nutritive antibiotics or chemotherapeutics in animal nutrition and in fulfillment of health claims in man and animals. Economic and environmental aspects will reduce the overall application of probiotics world-wide. Restrictions of use and controls of efficacy and safety are essential and must be implemented periodically.     

Probiotic Use in Horses – What is the Evidence for Their Clinical Efficacy?

The gastrointestinal microbiota is extremely important for human and animal health. Investigations into the composition of the microbiota and its therapeutic modification have received increasing interest in human and veterinary medicine. Probiotics are a way of modifying the microbiota and have been tested to prevent and treat diseases. Probiotics are proposed to exert their beneficial effects through various pathways. Production of antimicrobial compounds targeting intestinal pathogens, general immune stimulation, and colonization resistance are among these mechanisms. Despite widespread availability and use, scientific, peer‐reviewed evidence behind commercial probiotic formulations in horses is limited. Additionally, quality control of commercial over‐the‐counter products is not tightly regulated. Although promising in vitro results have been achieved, in vivo health benefits have been more difficult to prove. Whether the ambiguous results are caused by strain selection, dosage selection or true lack of efficacy remains to be answered. Although these limitations exist, probiotics are increasingly used because of their lack of severe adverse effects, ease of administration, and low cost. This review summarizes the current evidence for probiotic use in equine medicine. It aims to provide veterinarians with evidence‐based information on when and why probiotics are indicated for prevention or treatment of gastrointestinal disease in horses. The review also outlines the current state of knowledge on the equine microbiota and the potential of fecal microbial transplantation, as they relate to the topic of probiotics.     

Study and use of the probiotic Lactobacillus reuteri in pigs: a review

Probiotics are living microorganisms that provide a wide variety of health benefits to the host when ingested in adequate amounts. The bacterial strains most frequently used as probiotic agents are lactic acid bacteria, such as Lactobacillus reuteri, which is one of the few endogenous Lactobacillus species found in the gastrointestinal tract of vertebrates, including humans, rats, pigs and chickens. L. reuteri is one of the most well documented probiotic species and has been widely utilized as a probiotic in humans and animals for many years. Initially, L. reuteri was used in humans to reduce the incidence and the severity of diarrhea, prevent colic and necrotic enterocolitis, and maintain a functional mucosal barrier. As interest in alternatives to in-feed antibiotics has grown in recent years, some evidence has emerged that probiotics may promote growth, improve the efficiency of feed utilization, prevent diarrhea, and regulate the immune system in pigs. In this review, the characteristics of L. reuteri are described, in order to update the evidence on the efficacy of using L. reuteri in pigs.     

益生菌与功能发酵乳开发研究进展

近10 年来，随着先进研究方法的使用和研究内容的不断深入，对益生菌的认识取得重大进展，尤其是对益生菌促进健康作用的机理逐渐清晰，益生菌制备技术和产业化技术更加成熟，为新型发酵乳的开发提供了坚实的发展基础。本文对益生菌功能机理、遗传转化及代谢调控的研究现状和发展进行阐述，介绍分离自内蒙古马乳酒中的干酪乳杆菌LC2W和分离自发酵蔬菜中的植物乳杆菌ST-Ⅲ的益生特性、基因组及产胞外多糖特性，并进一步探讨基于植物乳杆菌增殖技术开发含高活菌数的纯植物乳杆菌发酵乳品，以期为益生菌和功能发酵乳的应用开发提供理论依据。发酵乳品是益生菌的良好载体，吸收现代益生菌科技成果，加强循证医学方法论证，基于生物活性效应分子及生物活性评价，增加临床研究数据，新型发酵乳将为人类健康做出更多贡献。     

Probiotics in animal nutrition: a review

The normal microflora colonizing the digestive tract in livestock species at five to six days after birth can be summarized as following: 400 to 500 different bacterial strains for a total count of 10(14) bacteriae. The role of the gut flora is multifarious: fermentation of feed compounds, induction of anatomical and physiological changes in the intestinal cell wall structure, increase in animal resistance against enteropathogenic bacteriae, etc. In the present review, the numerous hypotheses for explaining the beneficial effects of probiotics on zootechnical performances and also the healthy aspects for men and animals are summarized, and the possible mechanisms of bacterial attachment are enumerated. The different assumptions on mode of action and fixation mechanisms of probiotics in the gut are critically discussed, and the required characteristics of the ideal probiotic are listed. Finally, some zootechnical results obtained with lactic acid bacteria are shown for calves, piglets and broilers.     

Putative probiotic Lactobacillus spp. from porcine gastrointestinal tract inhibit transmissible gastroenteritis coronavirus and enteric bacterial pathogens

A total of 310 bacterial strains isolated from the porcine gastrointestinal tract were tested for their activity against transmissible gastroenteritis (TGE) coronavirus and other enteric pathogens. Based on activity, the strains Probio-38 and Probio-37 were selected as potential probiotics and identified as Lactobacillus plantarum Probio-38 and Lactobacillus salivarius Probio-37 respectively by 16S rRNA gene sequencing. Supernatants of these strains inhibited TGE coronavirus in vitro in ST cells, without any cytopathic effect even after 72 h of incubation. Both the strains exhibited high survival in synthetic gastric juice. The strains were resistant to 5% porcine bile and exhibited antimicrobial activity against all the 13 enteric bacterial pathogens tested. These strains also exhibited resistance to most of the antibiotics analyzed. The inhibition of transmissible gastroenteritis coronavirus and enteric bacterial pathogens as well as the bile tolerance, high survival in gastric juice, and the antibiotic resistance indicate that the two isolated bacterial strains are ideal probiotic candidates for animal application after proper in vivo experiments.     

益生菌健康宣称的调研分析

我国市场上将益生菌进行健康宣称的食品主要涉及调制乳粉、酸乳、乳酸菌饮料和益生菌粉。本次调研主要从产品中添加益生菌的种类、产品包装正面益生菌的健康宣称和产品包装背面益生菌的健康宣称三方面进行。同时对比我国与国外益生菌健康宣称的法规,针对国内益生菌健康宣称管理中的问题提出相关建议。结果表明：不同产品的益生菌添加种类不同,调制乳粉中添加的益生菌一般为1～2 种,而乳酸菌饮料、酸乳和益生菌粉中添加的益生菌大多在2 种以上,有些产品达到10 种以上;益生菌产品包装正面用语主要涉及“益生菌”、“活性”、“菌株名称”、     

Screening of the equine intestinal microflora for potential probiotic organisms

REASONS FOR PERFORMING STUDY: Probiotics have not been demonstrated to provide any beneficial health effects in horses, possibly because of improper selection of probiotic organisms. This study was designed to identify lactic acid bacteria of equine origin with predetermined beneficial properties which might make them useful as therapeutic probiotics. HYPOTHESIS: A small percentage of lactic acid bacteria that are native to the intestinal tract of horses possess properties that may be useful in the treatment and/or prevention of gastrointestinal disease in horses. METHODS: Faecal samples were collected from healthy mature horses and foals. Lactic acid bacteria were isolated and tested for the ability to grow in acid and bile environments, aerotolerance and in vitro inhibition of enteropathogens. One isolate that possessed these properties was administered orally to healthy mature horses and foals and gastrointestinal survival was assessed. RESULTS: Of the 47 tested organisms, 18 were deemed to be adequately acid- and bile-tolerant. All were aerotolerant. Four organisms markedly inhibited Salmonella spp. One isolate, Lactobacillus pentosus WE7, was subjectively superior and chosen for further study. It was also inhibitory against E. coli, moderately inhibitory against S. zooepidemicus and C. difficile and mildly inhibitory against C. perfringens. After oral administration, this isolate was recovered from the faeces of 8/9 (89%) foals and 7/8 (87.5%) mature horses. CONCLUSIONS: Lactobacillus pentosus WE7 possesses in vitro and in vivo properties that may be useful for the prevention and treatment of enteric disease in horses. POTENTIAL RELEVANCE: The beneficial in vitro and in vivo properties that L. pentosus WE7 possesses indicate that randomised, blinded, placebo-controlled efficacy studies are warranted.     

Health-beneficial effects of probiotics: Its mode of action

It is now widely recognized that probiotics have health-beneficial effects on humans and animals. Probiotics should survive in the intestinal tract to exert beneficial effects on the host's health. To keep a sufficient level of probiotic bacteria in the gastrointestinal tract, a shorter interval between doses may be required. Although adherence to the intestinal epithelial cell and mucus is not a universal property of probiotics, high ability to adhere to the intestinal surface might strongly interfere with infection of pathogenic bacteria and regulate the immune system. The administration of probiotic Lactobacillus stimulated indigenous Lactobacilli and the production of short-chain fatty acids. This alteration of the intestinal environment should contribute to maintain the host's health. The immunomodulatory effects of probiotics are related to important parts of their beneficial effects. Probiotics may modulate the intestinal immune response through the stimulation of certain cytokine and IgA secretion in intestinal mucosa. The health-beneficial effects, in particular the immunomodulation effect, of probiotics depend on the strain used. Differences in indigenous intestinal microflora significantly alter the magnitude of the effects of a probiotic. Specific probiotic strains suitable for each animal species and their life stage as well as each individual should be found.     

Porcine IPEC-J2 intestinal epithelial cells in microbiological investigations

IPEC-J2 cells are porcine intestinal columnar epithelial cells that were isolated from neonatal piglet mid-jejunum. This cell line forms polarized monolayers with high transepithelial electrical resistance when cultured on 0.4 μm pore-size filters. The cell line is unique in that it is derived from small intestinal tissue (compared to the common human colon-derived lines HT-29, T84, and Caco-2) and is not transformed (compared to the porcine small intestinal line, IPI-2I). Porcine intestinal epithelial cells more closely mimic human physiology than analogous rodent cell lines (e.g. IEC-6 or IEC-18), which is important in studies of zoonotic infections; in addition, they provide specificity to study porcine-derived infections. IPEC-J2 cells are increasingly being used in microbiological studies to examine the interactions of various animal and human pathogens, including Salmonella enterica and pathogenic Escherichia coli, with intestinal epithelial cells. The IPEC-J2 cell line has also been employed in some probiotic studies, in which the cells have been used as an initial screening tool for adhesiveness and anti-inflammatory properties of the potential probiotic microorganisms. The validity of these studies is not clear as follow-up studies to assess the efficacy of the probiotics in vivo have not been published to date. The aims of this review are to provide a comprehensive overview of the microbiological studies that have been conducted with IPEC-J2 cells and a reference guide of key cellular and immune markers that have been identified in this cell line that may prove to be useful in future studies.     

益生菌在马属动物中的应用及研究进展

益生菌作为一种活的微生物饲料补充剂,不仅能提高动物生产性能、增强免疫力,还有维持胃肠道菌群平衡、净化胃肠道环境的功能,在预防和治疗动物腹泻、调节宿主免疫功能等方面发挥着重要作用。益生菌具有安全性高、无残留、无耐药性等特点,可作为替抗产品。益生菌广泛应用于马属动物并取得良好的效果。综述了益生菌及相关活性物质的作用机理及马用益生菌产品存在的问题和应用前景,旨在为马用益生菌产品的开发利用提供参考。     

饲用益生菌产品评价研究进展

饲用益生菌是一种通过改善肠道菌群平衡而对动物产生有利影响的活的微生物,在饲料工业中有广泛的应用。作者针对益生菌在加工、使用过程中存在的问题,就饲用益生菌的评价现状及研究进展做了简要的评述,并根据人畜消化生理的异同,借鉴《食品益生菌评价指南》,初步提出饲用益生菌产品评价指标。     

Evaluation of Lactobacillus pentosus WE7 for prevention of diarrhea in neonatal foals

OBJECTIVE: To evaluate the efficacy of Lactobacillus pentosus WE7, an equine-origin organism with potentially beneficial in vitro properties, as a probiotic for prevention of neonatal diarrhea in foals. DESIGN: Randomized controlled clinical trial. ANIMALS: 153 foals. PROCEDURE: Foals were enrolled at 24 to 48 hours of age and randomly assigned to treatment or control groups. The treatment group received approximately 2 x 10(11) CFU of freeze-dried L. pentosus WE7 orally once daily for 7 days, whereas the control group received a placebo. Clinical monitoring was performed for 14 days. RESULTS: Probiotic administration was significantly associated with development of signs of depression, anorexia, and colic and the need for veterinary examination and treatment. Probiotic-treated foals also had more days of diarrhea, compared with the control group, although not significantly. In a multivariate model, probiotic administration was significantly associated with development of diarrhea and diarrhea plus additional clinical abnormalities. CONCLUSIONS AND CLINICAL RELEVANCE: Administration of L. pentosus WE7 did not prevent diarrhea; rather, it was associated with development of diarrhea and, most importantly, development of diarrhea plus additional clinical abnormalities and the need for veterinary intervention. The promising in vitro properties of L. pentosus WE7 were not evident in vivo. Results raise concern about the variety of untested probiotic products that are commercially available. Safety and efficacy testing needs to be performed for all potential equine probiotics.     

功能性益生菌菌种的选育和益生性研究

本研究从新疆、内蒙古分离到30株乳酸杆菌和乳酸球菌,经过鉴定并选择发酵性能较好的菌种进行进一步的研究。经过对益生乳杆菌人工胃肠液耐受性、胆盐耐受性、胆固醇降解能力等益生特性的研究,得出植物乳杆菌BN-3、BN-5、罗伊氏乳杆菌AL3、鼠李糖乳杆菌AL20具有较好的益生特性。     

可食用益生菌在肾健康维护中的活性与机制研究进展

肾健康问题已经是重要公共卫生问题,也是容易被忽视的问题,现有研究表明,通过益生菌调整肠道菌群是肾健康维护的新型干预路径.然而,可食用益生菌在肾健康维护中的作用目前还缺乏关注.本文围绕可食用益生菌应用于肾健康相关的慢性肾炎、血透析、高尿酸及高草酸相关的基础研究进行调研,并对相关疾病干预涉及的可食用益生菌种类、作用效果和作...     

乳酸菌在家禽生产中的应用

乳酸菌是人、畜禽胃肠道以及一些食物中的重要菌群之一,广泛用于食品、饲料和医药等领域,是一类具备多种益生功能的微生物。以乳酸菌为代表的微生态制剂,具有改善机体生长性能、促进机体免疫系统发育防止发生疾病、维持机体胃肠道微生态平衡、提升抗氧化能力、促进机体对营养物质的消化吸收等作用,一直是各大饲料企业以及家禽养殖业关注的焦点。文章主要对乳酸菌的分类、生理功能及不同乳酸菌在家禽生产中的应用展开叙述。     

体外发酵产气技术在饲料营养价值评定中的应用

体外产气技术是反刍动物饲料营养价值评定中常用的方法之一。文章比较了体内法、瘤胃尼龙袋法和体外法在反刍动物营养价值评定中的特点,并着重从体外产气技术的技术细节、测定装置和应用范围进行了全面的论述,为该技术的标准化和自动化提供参考。