Effects of lactic acid bacteria and smectite after aflatoxin B1 challenge on the growth performance,nutrient digestibility and blood parameters of broilers

This study aimed to investigate the effect of lactic acid bacteria (LAB) and smectite on the growth performance, nutrient digestibility and blood parameters of broilers that were fed diets contaminated with aflatoxin B₁ (AFB₁). A total of 480 newly hatched male Arbor Acres broilers were randomly allocated into four groups with six replicates of 20 chicks each. The broilers were fed diets with the AFB₁ (40 μg/kg) challenge or without (control) it and supplemented with smectite (3.0 g/kg) or LAB (4.0 × 10¹⁰ CFU/kg) based on the AFB₁ diet. The trial lasted for 42 days. The results showed that during days 1–42 of AFB₁ challenge, the feed intake (FI) and body weight gain (BWG) were depressed (p < .05). The inclusion of LAB and smectite increased (p < .05) the BWG by 71.58 and 41.89 g/bird, respectively, which reached the level of the control diet (p ≥ .05), but there were no differences (p ≥ .05) in performance between LAB and smectite. LAB and smectite also increased (p < .05) the apparent total tract digestibility of the crude protein. Regarding the blood parameters, AFB₁ decreased (p < .05) the levels of red blood cell count, haematocrit, mean corpuscular volume, haemoglobin, albumin and total protein. In the meantime, the AFB₁ increased (p < .05) leucocyte counts, urea nitrogen, cholesterol, total bilirubin, creatinine, glutamic‐pyruvic transaminase, glutamic oxaloacetic transaminase and alkaline phosphatase. By contrast, LAB and smectite affected (p < .05) these parameters in the opposite direction. It can be concluded that after the AFB₁ challenge, LAB and smectite have similar effects on the growth and health of the broilers, suggesting that LAB could be an alternative against AFB₁ in commercial animal feeds.     

乳酸菌清除霉菌毒素作用及其机制

霉菌毒素广泛存在于食品和饲料当中,严重地危害人类和动物的健康。如何有效清除霉菌毒素一直是重要的研究课题。乳酸菌具有多种生物学功能,近年来研究表明,乳酸菌具有清除霉菌毒素能力。为了更好地理解乳酸菌清除霉菌毒素的作用,文章就近年来乳酸菌对霉菌毒素的清除机制加以综述。     

Identification and characterization of lactic acid bacteria isolated from mixed pasture of timothy and orchardgrass,and its badly preserved silage

In order to understand the relationship between lactic acid bacteria (LAB) species and silage fermentation, a total of 65 LAB strains isolated from mixed pasture of timothy (Phleum pratense L.) and orchardgrass (Dactylis glomerata L.), and its badly preserved silages were subjected to phenotypic and genetic analysis. According to these analyses, the isolates were divided into 13 groups, including Enterococcus gallinarum, Lactobacillus acidipiscis, L. coryniformis subsp. coryniformis, L. coryniformis subsp. torquens, L. curvatus, L. paraplantarum, L. plantarum subsp. argentoratensis, L. plantarum subsp. plantarum, L. sakei subsp. carnosus, Lactococcus garvieae, Lactococcus lactis subsp. cremoris, Leuconostoc pseudomesenteroides, Pediococcus acidilactici, Pediococcus pentosaceus, Weissella hellenica, Weissella paramesenteroides and Carnobacterium divergens. This is the first report to document that C. divergens, L. acidipiscis, L. sakei subsp. carnosus, L. garvieae, phenotypically novel L. lactis subsp. cremoris, E. gallinarum and W. hellenica are present in vegetative forage crops. L. plantarum group strains were most frequently isolated from the badly preserved silages. Some isolates showed a wide range of growth preferences for carbohydrate utilization, optimal growth pH and temperature in vitro, indicating that they have a high growth potential. These results are useful in understanding the diversity of LAB associated with decayed silage of timothy and orchardgrass.     

16S ribosomal DNA analysis and characterization of lactic acid bacteria associated with traditional Tibetan Qula cheese made from yak milk

Twenty strains of lactic acid bacteria were isolated from six traditional Tibetan Qula cheese made from yak which were collected from northwest China, including Tibet, Qinghai and Gansu province. These isolates were subjected to phenotypic and genetic analyses. All isolates were Gram‐positive and catalase‐negative cocci that produced gas from glucose and formed D(–) isomer of lactate. Most isolates were able to grow in de Man, Rogosa and Sharpe (MRS) broth at pH values 3.0–9.0 and in 6.5% NaCl (w/v). According to analytical profile index 50 carbohydrates (API 50 CH) fermentation patterns of amygdalin and arabinose, these isolates were divided into three groups (A to C). On the basis of the phylogenetic trees of 16S ribosomal DNA (rDNA) sequence, the strains in all groups were placed in the cluster making up the genus Leuconostoc, which showed that all strains should belong to Leuconostoc species. Strains in Group A and Group B exhibited similarity of 16S rDNA sequence of over 99% to Leuconostoc mesenteroides, indicating that they each comprised a single species. Strains in group C were assigned to the Leuconostoc pseudomesenteroides and their 16S rDNA sequence showed a similarity of over 99%. This study demonstrated that Leuconostoc was the dominant member among lactic acid bacteria in Qula cheese.     

云南传统乳产品中乳酸菌的菌群分析

对采集于云南传统乳制品中乳酸菌的种类及优势菌群的分布情况进行了研究.从采集的8份样品中共分离获得73株乳酸菌,分属于5个属的13个种.对乳酸菌优势菌群分析,发现肠球菌、乳球菌和同型发酵乳杆菌为主要菌群,分别占乳酸菌总数的20%、27%和20%.     

Palatability and physicochemical properties of sausages prepared via lactic acid fermentation and drying at low temperature

The present study aimed to assess the palatability of sausages undergoing low‐temperature fermentation and drying process. Lactobacillus sakei D‐1001 or Lactobacillus salivarius A001 were used as starter cultures for fermentation, and the following properties of the sausages were investigated: colony‐forming units of lactic acid bacteria; concentrations of lactic acid, protein, peptides, and free amino acids; distribution of protein; composition of free amino acids; and physical properties and taste. Alterations in the composition of proteins, peptides, and free amino acids as well as in various physical properties were caused by fermentation by lactic acid bacteria. A sensory test indicated that the palatability of the fermented sausages was greater than that of the non‐fermented sausages, particularly in terms of hardness and juiciness. This was considered to be due to protein degeneration and changes in the physical properties of the sausages as a result of fermentation by lactic acid bacteria. However, the taste of the fermented sausages was sourer than that of the non‐fermented sausages, and therefore, inferior. Our study revealed that the palatability of the sausages in terms of hardness and juiciness were increased by low‐temperature fermentation by lactic acid bacteria and the drying process.     

对禾草源同型发酵和/或异型发酵乳酸菌发酵无芒雀麦青贮有氧稳定性的评价

本研究旨在评价禾草源同型发酵和/或异型发酵乳酸菌发酵对无芒雀麦青贮有氧稳定性的作用效果。将从禾本科牧草上分离筛选出的2株同型发酵植物乳杆菌(Lactobacillus plantarum)和蒙氏肠球菌(Enterococcus mundtti)及1株异型发酵布氏乳杆菌(Lactobacillus buchneri)制成发酵剂。选用开花期无芒雀麦,刈割切段至2~3 cm,根据所喷发酵剂菌株的不同,分为4个处理:1)对照处理(con处理),喷洒无菌去离子水;2)异型发酵处理(he处理),喷洒布氏乳杆菌液;3)同型发酵处理(ho处理),喷洒植物乳杆菌和蒙氏肠球菌混合液(2种菌1∶1混合);4)同型发酵+异型发酵处理(he+ho处理),喷洒布氏乳杆菌、植物乳杆菌和蒙氏肠球菌混合液(3种菌1∶1∶1混合)。各处理喷洒量均为10 m L/kg鲜牧草,喷洒总菌数约为5×105CFU/g鲜牧草,每个处理设5个重复。实验室常温发酵60 d后开封,通过测定无芒雀麦青贮营养成分、菌群数量及中心温度变化评价其有氧稳定性。结果显示:经60 d青贮发酵后,4个处理的p H均较青贮前显著降低(P0.05),所有处理发酵效果均较好(p H≤4.3),尤以ho处理效果最优。而在有氧暴露试验期间,con和ho处理的p H迅速增加,至第8天时达到7以上;he+ho处理p H增加幅度略低于con和ho处理,其p H至第8天时为6.3,显著低于con和ho处理(P0.05);he处理p H增加非常缓慢,至第8天时仅为4.4,显著低于其他处理(P0.05)。经60 d青贮发酵后,可溶性碳水化合物(WSC)含量以ho处理最高,显著高于其他处理(P0.05);con处理最低,显著低于其他处理(P0.05)。而在有氧暴露试验期间,WSC含量在ho处理中迅速降低,在he处理中缓慢降低;至第8天时,WSC含量以he处理最高,he+ho处理显著低于前者(P0.05),而con和ho处理则显著低于he和he+ho处理(P0.05)。青贮发酵后霉菌数量明显受到抑制,且以单独添加异型发酵乳酸菌和混合添加同型发酵和异型发酵乳酸菌时抑制效果较优,这2个处理均未检出霉菌,且he+ho处理在有氧暴露至第5天时仍未检出霉菌,he处理在有氧暴露至第8天时仍未检出霉菌。在有氧暴露第3和5天时,he和he+ho处理酵母菌数量显著低于con、ho处理(P0.05)。在有氧暴露第8天时,he+ho与he处理的乳酸、乙酸浓度均显著高于另2个处理(P0.05)。he、he+ho、con和ho处理的有氧稳定性依次降低,分别为194、126、62和58 h。综合评价结果得出:布氏乳杆菌单独或与同型发酵乳酸菌蒙氏肠球菌和植物乳杆菌联合接种可有效抑制无芒雀麦青贮的有氧腐败,保证无芒雀麦青贮有氧暴露期间品质的稳定,且前者更有效;接种同型发酵乳酸菌蒙氏肠球菌和植物乳杆菌未能在提高无芒雀麦青贮有氧稳定性方面表现出积极的作用。     

Selection of useful probiotic lactic acid bacteria from the Lactobacillus acidophilus group and their applications to functional foods

In the present review, a new mass screening system for selecting probiotic strains from Lactobacillus (L) acidophilus group lactic acid bacteria (LAB) with strong adhesion to the human intestinal tract is described. Characteristics of antimicrobial peptides (bacteriocin), lactose‐hydrolyzing enzymes and immunostimulative oligo DNA motifs in L. gasseri strains are also described. Finally, the use of L. acidophilus LAB, selected by our screening method, that have strong adhesion to the human colonic mucosa in functional yogurt products is described. Adhesiveness to the human intestine is one of the most important characteristics of probiotic LAB. A new screening system that involves a combination of three methods is proposed: rat colonic mucin (RCM)‐micro plate assay, Carnoy's histochemical staining method and carbohydrate probe binding assay. By using an RCM‐coated poly‐vinylidene‐diflouride membrane that mimics the human colonic mucous layer, a new lectin was isolated and its structure was clarified by gene cloning. Furthermore, the structures and functions of a new cyclic bacteriocin (gassericin A), new lactose‐hydrolyzing enzymes, and new immunostimulating oligo DNA motifs from Lactobacillus gasseri (B₁ subgroup) were clarified. A new functional yogurt ‘Fit down’ is proposed, that is fermented by an adhesive strain of L. acidophilus LA67 selected by our screening and contains antihypertensive peptides derived from whey proteins by protease digestion. In the future, superior functional foods containing more effective probiotic LAB are expected to be developed by the use of the proposed mass screening system.     

Genotypic and phenotypic characterization of lactic acid bacteria isolated from Italian ryegrass silage

Twenty-three lactic acid bacteria (LAB) isolated from three cultivars (Akiaoba, Nagahahikari and Tachiwase) of Italian ryegrass (Lolium multiflorum Lam.) silage were precisely characterized by a combination of phenotypic tests, genotypic 16S ribosomal DNA sequencing and rapid PCR-based analyses, focusing on their useful phenotypes for silage preparation as inoculants. We successfully identified both at the species and subspecies levels: phenotypically novel Lactococcus lactis subsp. lactis, Lactobacillus brevis, Lactobacillus coryniformis subsp. torquens, Lactobacillus curvatus, Lactobacillus plantarum subsp. plantarum, Lactobacillus sakei subsp. carnosus, Leuconostoc mesenteroides subsp. dextranicum and Pediococcus parvulus. This is the first report to elucidate the presence of Lactobacillus coryniformis ssp. torquens and Leuconostoc mesenteroides subsp. dextranicum in Italian ryegrass silages. Physiological and biochemical tests revealed that phenotypic characteristics are different among the different strains of the same species and subspecies, and that the isolates show unique and diverse phenotypes related to fermentation factors, such as available carbohydrates, optimal growth pH and temperature. These results suggest that, for various well-preserved silage preparations, the isolates may be useful in producing novel inoculants corresponding to their optimally climatic and ecological niches.     

乳酸菌发酵扶正解毒散复合制剂的制备及其在肉鸡上的应用研究

本研究旨在制备一种乳酸菌发酵的扶正解毒散复合制剂,并探索其对肉鸡生长性能和免疫器官发育的影响。首先,利用单因子试验法将不同比例的黄芪、板蓝根、淫羊藿及复合中药提取液添加至乳酸菌的培养基中,测定中药对乳酸菌生长的影响;其次,利用正交试验对复合药液培养基进行优化,确定最佳的培养基配方;最后,选取1日龄健康AA肉仔鸡450只,随机分为3组,每组3个重复:对照组(抗生素)、试验Ⅰ组(发酵制剂)、试验Ⅱ组(未发酵制剂),分别在试验的第21、42天测定肉鸡的生长及免疫器官指标。试验结果显示,黄芪、板蓝根、淫羊藿及复合药液组的最高活菌数与对照组相比分别提高了261.64%、505.48%、156.16%、564.38%。通过正交试验得到复合发酵液最适生产培养基配方:复合药液100 mL、红糖2.0 g、蛋白胨0.3 g、豆粕2.0 g,发酵液活菌数为7.13×10⁹ CFU/mL。肉鸡生长试验结果显示,1~42日龄阶段,Ⅰ、Ⅱ组的平均日增重分别比对照组提高了17.84%和8.22% (P<0.05),料肉比分别比对照组降低了10.88%和4.66% (P<0.05),平均日采食量明显高于对照组 (P<0.05);21、42日龄时,Ⅰ、Ⅱ组脾指数分别比对照组提高了11.04%和13.58% (P<0.05)、7.79%和7.41% (P<0.05),法氏囊指数分别比对照组提高了31.30%和26.23% (P<0.05)、15.65%和11.48% (P<0.05)。以上试验结果表明中药提取液能促进乳酸菌的增殖,复合发酵液能够提高肉鸡的生长性能并促进免疫器官的发育。     

乳酸菌重组表达抗菌肽研究进展

本文对乳酸菌表达系统的载体、筛选标记及表达形式进行概述,阐述了抗菌肽在乳酸菌中重组表达的研究进展以及乳酸菌表达抗菌肽的优势和存在的问题。     

Studies on Preparation and Application of Fuzhengjiedusan Compound Preparation Fermentated by Lactic Acid Bacteria in Broilers

This study was aimed toproduce compound preparations of Fuzhengjiedusan fermented with Lactic acid bacteria and analyze its effectson growth performancesand immune organ growth of broilers. Firstly, to identify the influence of traditional Chinese medicine on the growth of Lactic acid bacteria, the medium for the growth of Lactic acid bacteria was filled with different proportions of Astragalusmembranaceus, Radix Isatidis, Epimedium and compound Chinese herbal medicine extracts by single factor test. Secondly, to confirm the optimum medium, orthogonal test was used to optimize the culture medium of compound medicine liquid. Finally, 450 one-day-old healthy AA broilers were randomized into three groups, each group had 3 repeats: control group (antibiotics), group Ⅰ(fermented preparation), group Ⅱ (unfermented preparation). The growth and immune organ index were determined in 21 and 42 days after administration. The results showed that the highest viable count of Astragalusmembranaceus, Radix Isatidis, Epimedium and compound medicine liquid groups were increased by 261.64%, 505.48%, 156.16% and 564.38%, respectively, compared with control group. The optimum medium composition was compound medicine liquid 100 mL, brown sugar 2.0 g, peptone 0.3 g, soybean meal 2.0 g. The viable count of fermentation liquid was 7.13×10⁹ CFU/mL.The production results showed that in period of 1~42 days, the daily weight gain of group Ⅰ and Ⅱ were separately increased by 17.84% and 8.22% (P<0.05) than that of the control. What's more, the feed conversion rate was reduced by 10.88% (P<0.05). The average daily feed intake was significantly higher than control group (P<0.05). At 21 and 42 days, the spleen index of group Ⅰ were improved by 11.04% and 13.58% (P<0.05), respectively. The bursal index were raised by 31.30% and 26.23% (P<0.05), respectively. However, the spleen index of group Ⅱ were improved by 7.79% and 7.41% (P<0.05), respectively. The bursal index were raised by 15.65% and 11.48% (P<0.05), respectively. On the basis of the above results, it could be suggested that Chinese herbal medicine extracts can promote lactic acid bacteria proliferation and compound fermentation liquid can improve growth performance in broilers, as well as immune organ development.     

添加丙酸和乳酸菌对杂交狼尾草青贮发酵品质的影响

研究丙酸、乳酸菌和两者组合不同添加水平对杂交狼尾草（Pennisetum americanum×P.purpereum）青贮发酵品质的影响。试验设对照(无添加)、乳酸菌(LAB)、0.2%丙酸、0.3%丙酸、0.4%丙酸、0.5%丙酸、0.2%丙酸+乳酸菌、0.3%丙酸+乳酸菌、0.4%丙酸+乳酸菌、0.5%丙酸+乳酸菌（占鲜质量比例）处理,青贮45 d后开袋分析。结果表明,添加丙酸可以有效改善杂交狼尾草的青贮品质,最佳添加量为0.3%;添加乳酸菌有助于提高杂交狼尾草的有氧稳定性。     

乳酸菌产生的新型抗菌物质——苯乳酸的抑菌性质及作用机理研究

苯乳酸是微生物产生的一种新型天然抗菌物质,已成为乳酸菌抑菌能力的有效标志之一。与乳酸菌产生的细菌素(如乳链球菌素)相比,苯乳酸是小分子物质、抑菌谱宽、稳定性高。作为一种新型生物防腐剂,苯乳酸在乳品工业中具有广阔的应用前景。研究表明苯乳酸对多种食源性致病菌如致病性大肠杆菌、单增李斯特菌和金黄色葡萄球菌,腐败菌包括产毒素的丝状真菌如赭曲霉、疣孢青霉和桔青霉,有很强的抑菌作用。目前的研究表明苯乳酸的作用靶位之一是微生物的细胞壁,但阐明苯乳酸的作用机理需要采用相关技术进一步研究其他作用位点。本文对苯乳酸的抑菌活性、作用机理等研究进展进行综述,并展望了今后的研究方向。     

乳酸菌青贮添加剂的研究与应用

很多添加剂被用来促进青贮发酵过程,其中研究最广泛的就是乳酸菌青贮添加剂。本文主要阐述了在青贮中加入乳酸菌添加剂可以促进作物的发酵速度,并且改善家畜的生产性能。     

内生真菌提高禾草耐重金属胁迫的研究进展

随着全球工业化和地球化学活动的日益频繁, 地表和地下水中重金属污染已经成为一个严重的环境问题。植物通过体内存在的内生真菌提高对重金属的耐受性进行修复。本研究围绕重金属胁迫对禾草的影响、禾草对重金属胁迫的响应以及内生真菌提高禾草耐重金属的机理等方面对内生真菌提高禾草耐重金属胁迫的研究进展方面进行了综述, 以期为禾草耐重金属胁迫品种的选育和植物修复机制的研究, 提供新的方法手段和理论基础。     

单一和复合乳酸菌添加水平对苜蓿青贮营养品质及蛋白组分的影响

以第1茬现蕾期刈割的紫花苜蓿(Medicago sativa)为原料(干物质含量为42.56%),结合康奈尔碳水化合物-蛋白质体系(CNCPS),研究不同浓度的单一和复合乳酸菌对苜蓿青贮品质及蛋白组分的影响。发酵40d后取样分析其青贮发酵品质和营养成分,结果表明,所有乳酸菌处理组都能显著降低苜蓿青贮饲料的氨态氮含量(P0.05),并且均未检测到丁酸;添加量为1×10~7 cfu·g~(-1)的单一乳酸菌处理组氨态氮含量最低,乳酸含量最高;除添加量为1×10~5cfu·g~(-1)的复合菌处理组外,其它乳酸菌处理组的pH均显著低于无添加剂处理组(P0.05);乳酸菌处理后粗蛋白含量无显著变化(P0.05);复合菌处理组的非蛋白氮含量显著低于无添加剂处理组(P0.05);单一和复合乳酸菌都能降低蛋白组分中的非蛋白氮(PA)含量,增加快速降解真蛋白质(PB1)和中速降解真蛋白质(PB2)含量,复合菌处理的真蛋白质(PB)含量显著高于对照的(P0.05)。综上所述,高浓度的单一植物乳杆菌(Lactobacillus plantarum)处理的苜蓿青贮发酵品质最好,复合菌能保护更多的真蛋白不被降解。     

Fermentation quality and chemical composition of shrub silage treated with lactic acid bacteria inoculants and cellulase additives

Effects of lactic acid bacteria (LAB) inoculants and cellulase additives on fermentation quality and chemical compositions of shrub silages were studied by using a small‐scale fermentation system. Two LAB inoculants of Qingbao (Lactobacillus plantarum, Pediococcus acidilacticii, Lactobacillus casei and Clostridium phage) and Caihe (Lactobacillus plantarum, Lactobacillus brevis and Pediococcus acidilactici) and a commercial cellulase made from Trichoderma reesei were used as additives for intermediate pea‐shrub, rush bushclover, arborescent ceratoides and shrubby silage preparation. The crude protein, neutral detergent fiber and water‐soluble carbohydrate contents of the four shrub materials were 10.1–14.2, 62.6–67.2 and 1.9–3.5% on a dry matter basis, respectively. All shrub silages had pH 3.40–4.43, ammonia‐N 0.1–0.2% g/kg and lactic acid 1.3–2.9% on a fresh matter basis. The silage quality of LAB‐inoculated silages did not have a greater effect than control silages, except shrubby silage preparation. Silages treated with the cellulase, the pH of rush bushclover and shrubby sweetvetch silage were significantly (P < 0.05) lower and the lactic acid content were significantly (P < 0.05) higher than the control silages. The results confirmed that shrub contained a relatively high content of crude protein; its silages can be preserved in good quality, and they are new potential resources for livestock feed.     

Characterization and application of lactic acid bacteria for tropical silage preparation

Strains TH 14, TH 21 and TH 64 were isolated from tropical silages, namely corn stover, sugar cane top and rice straw, respectively, prepared in Thailand. These strains were selected by low pH growth range and high lactic acid‐producing ability, similar to some commercial inoculants. Based on the analysis of 16S ribosomal RNA gene sequence and DNA‐DNA relatedness, strain TH 14 was identified as Lactobacillus casei, and strains TH 21 and TH 64 were identified as L. plantarum. Strains TH 14, TH 21, TH 64 and two commercial inoculants, CH (L. plantarum) and SN (L. rhamnosus), were used as additives to fresh and wilted purple Guinea and sorghum silages prepared using a small‐scale fermentation method. The number of epiphytic lactic acid bacteria (LAB) in the forages before ensilage was relatively low but the numbers of coliform and aerobic bacteria were higher. Sorghum silages at 30 days of fermentation were all well preserved with low pH (3.56) and high lactic acid production (72.86 g/kg dry matter). Purple Guinea silage inoculated with LAB exhibited reduced count levels of aerobic and coliform bacteria, lower pH, butyric acid and ammonia nitrogen and increased lactic acid concentration, compared with the control. Strain TH 14 more effectively improved lactic acid production compared with inoculants and other strains. © 2016 Japanese Society of Animal Science     

香蕉茎叶中乳酸菌的分离和鉴定

从海南热带地区的香蕉茎叶中分离出3株高产酸菌,利用经典分类法即在形态学、生理生化特性等方面对其进行了细菌学鉴定,结果表明,3株菌分别为植物乳杆菌(Lactobacillus planetarium)、短乳杆菌(Lactobacillus brevis)和乳酸片球菌(P.pentosaceus)。对这3株菌产酸性能的测试结果表明,植物乳杆菌HN1和短乳杆菌HN2产酸速率大于乳酸片球菌HN3。