发酵鱼制品中乳酸菌的筛选鉴定及其初步应用

自传统发酵鱼制品中分离得到98株乳酸菌,通过发酵适应性和发酵特性等试验筛选出2株优良乳酸菌Z40和Y33,其具有良好的生长特性、产酸性能和耐盐性;经16S rDNA序列分析鉴定,乳酸菌Z40和Y33均为植物乳杆菌。将乳酸菌Z40应用于鱼糜发酵,结果表明该菌可快速降低鱼糜体系的pH值,显著抑制挥发性盐基氮的形成。     

自然发酵肉制品中乳酸菌的分离及特性研究

以MRS和改良TJA(改良番茄汁琼脂培养基)培养基为基础培养基,从自然发酵肉品中分离得到8个菌株,其中4株为乳酸菌,3株为杆菌,1株为球状菌。进一步测试表明:4株乳酸菌在30℃培养时,生长最好;在pH3.5～4.0时生长良好;4株乳酸菌在10%NaCl的环境中均能生长,其中以2%～6%的NaCl浓度内生长最适宜;属兼性厌氧菌;对大肠杆菌产生一定的抑制作用。分离出的4株乳酸菌均适宜作为发酵肉制品的发酵剂。     

发酵米浆中高发酵性能酵母菌和乳酸菌的筛选和鉴定

为了实现传统米发糕的规范化工业生产,从传统米发糕的发酵米浆中筛选发酵性能较好的菌株,并进行种属鉴定。首先从发酵米浆中分离出40 株酵母菌的疑似菌和30 株乳酸菌的疑似菌。经杜氏小管产气、发酵液特性和生长曲线分析三级筛选,最终得到1 株高发酵性能的酵母菌。经革兰氏染色等初筛、pH 值和生长曲线分析,最终得到1 株高发酵性能的乳酸菌。最后对两株菌株进行菌落形态观察和生理生化种属鉴定,确定分别为卡斯特酒香酵母和植物乳杆菌。     

发酵风干肠中乳酸菌的发酵性能

对从自然发酵风干肠中分离的6 株乳酸菌的发酵性能进行评价,主要包括测定菌株的生长曲线、产酸能力、对NaCl及NaNO2的耐受能力,同时通过吲哚实验和抗生素敏感性测定对菌株的安全性进行初步评价。结果表明：6 株乳酸菌生长趋势接近,均在8 h左右进入生长稳定期,pH值在0～8 h下降最快,清酒乳杆菌和发酵乳杆菌的产酸能力更强;所有菌株均可在6 g/100 mL NaCl和0.015 g/100 mL NaNO2条件下生长,植物乳杆菌和弯曲乳杆菌的NaCl耐受能力最优,清酒乳杆菌和发酵乳杆菌则对NaNO2具有最强的耐受能力;吲哚实验中6 株乳酸菌的反应结果均为阴性,对实验所选抗生素无耐药性,说明6 株乳酸菌具有较好的发酵性能和安全性,可作为功能性发酵剂用于发酵肉制品的生产。     

乳酸菌在发酵鱼制品加工中的应用研究概述

乳酸菌是能发酵乳糖或葡萄糖产生乳酸的一类革兰氏阳性球菌或杆菌,无芽孢、厌氧或兼性厌氧,在食品等行业广泛应用,可以改善食品的安全性、感官和营养特性。乳酸菌发酵应用在鱼制品中可以有效延长贮藏期,改善发酵鱼制品的品质。该文总结分析了现有的相关文献,介绍乳酸菌的分类及生物学功能,分析乳酸菌发酵对发酵鱼制品的保藏与品质的影响,对比传统发酵与人工接种发酵鱼制品,以期为乳酸菌发酵鱼制品深加工的研究与应用提供参考。     

Screening and Application from Fermented of Lactic Acid Bacteria Fish Products

自传统发酵鱼制品中分离得到98株乳酸菌,通过发酵适应性和发酵特性等试验筛选出2株优良乳酸菌Z40和Y33,其具有良好的生长特性、产酸性能和耐盐性;经16SrDNA序列分析鉴定,乳酸茵Z40和Y33均为植物乳杆菌。将乳酸菌Z40应用于鱼糜发酵,结果表明该菌可快速降低鱼糜体系的pH值,显著抑制挥发性盐基氮的形成。     

传统东北酸菜中优良发酵性能乳酸菌的筛选及应用

以传统东北酸菜为研究对象,从中分离筛选高产酸乳酸菌,通过探究乳酸菌的低温生长性能、耐盐特性及抑菌能力,从中获得优良发酵性能的乳酸菌,其经分子生物学鉴定后应用于发酵酸菜。结果表明,从传统东北酸菜分离筛选获得192株乳酸菌,其中5株为高产酸乳酸菌,均被鉴定为植物乳杆菌（Lactobacillus plantarum）;5株高产酸乳酸菌中植物乳杆菌DBC3和DBC33兼有多种优良性能,且在15 ℃条件下,接种量为3%的植物乳杆菌DBC33发酵得到的酸菜感官评分最高,显著缩短了发酵周期（P＜0.05）,为东北酸菜的标准化生产和酸菜发酵剂的制备提供了参考依据。     

低盐发酵香肠用优良菌株的分离筛选及鉴定

从10 种我国自然发酵制品中分离纯化得到52 株乳酸菌及56 株过氧化氢酶阳性球菌,首先通过菌株耐受性、发酵特性等指标对菌株进行初步筛选,然后利用抑菌特性和风味特性对菌株进行复筛,选择可能改善低盐发酵香肠安全性和风味的优良菌株,最终筛选得到3 株葡萄球菌Z9、L2和R2,以及4 株乳酸菌P6、P12、X和SN1-3。经由形态学特征、生理生化特征及16S rDNA序列分析对筛选得到的菌株进行鉴定,结果表明,葡萄球菌菌株Z9、L2为腐生葡萄球菌,R2为肉葡萄球菌;乳酸菌菌株P6、P12均为植物乳杆菌,X为干酪乳杆菌,SN1-3为戊糖片球菌。最后测定菌株的生长特性及产酸能力,研究菌株间的拮抗特性,最终筛选得到戊糖片球菌SN1-3与肉葡萄球菌R2作为制作发酵香肠的复配菌株。     

自然发酵肉制品中乳酸菌的筛选

为获得适合肉制品发酵的优良乳酸菌菌株,对来源于自然发酵肉制品中的25株乳酸菌进行筛选,其中19株在6 g/100 mL NaCl和150 mg/kg NaNO_2液体培养基中生长较好,结合其他发酵特性实验,13株乳酸菌符合肉制品发酵剂标准,挑选其中3株进行产酸能力、生长曲线及不同温度条件下生长情况的研究。结果表明:3株菌均具有较强的增殖及产酸能力;培养12 h时菌株即进入稳定生长期,培养24 h时培养基pH值降到4左右;在肉制品的发酵温度范围内能较好地生长。经形态学和16S rRNA序列分析,3株菌均为弯曲乳杆菌,它们均可作为潜在的开发肉制品发酵剂的出发菌株。     

发酵鱼酱酸产GABA乳酸菌的分离筛选及发酵特性

从传统发酵鱼酱酸中筛选出产γ-氨基丁酸（γ-aminobutyric acid，GABA）的乳酸菌并分析其菌株发酵特性。通过薄层色谱法定性、Berthelot比色法定量获得产GABA菌株，并进行耐酸、耐胆盐、氨基酸脱羧酶活性、抑菌性、生长曲线及pH值、产酸速率等菌株发酵特性分析。结果表明：从分离自鱼酱酸不同发酵阶段的387 株乳酸菌中，获得15 株典型产GABA菌株，包括2 株食窦魏斯氏菌（Weissella cibaria）、1 株熊蜂魏斯氏菌（Weissella bombi）、11 株植物乳杆菌（Lactobacillus plantarum）和1 株戊糖乳杆菌（Lactobacillus pentosus）。15 株乳酸菌产GABA能力及发酵特性在主成分分析图上差异显著，其中食窦魏斯氏菌Y113产GABA量为0.239 mg/mL，高于其他菌株；植物乳杆菌Y279和Y64展现出较好的耐酸性、耐胆盐性、抑菌性、无氨基酸脱羧酶活性、生长速率及产酸速率快的特点。鉴于其优良的发酵特性、益生特性及产GABA能力，菌株Y279可作为鱼酱酸工业化、标准化生产的潜在优良菌株。     

风干武昌鱼中乳酸菌的分离鉴定及发酵性能研究

为获得具备优良性能的乳酸菌发酵剂,以我国极具特色的发酵肉制品—风干武昌鱼为基础原料,对传统风干武昌鱼在自然发酵过程中不同时期的乳酸菌进行分离鉴定与发酵性能测定。研究通过平板分离、菌落与菌体形态观察、糖醇发酵试验及生理生化试验鉴定所分离乳酸菌。结果表明所分离乳酸菌分属于7个种属:嗜盐四联球菌(Tetragenococcus halophilus)、弯曲乳杆菌(Lactobacillus curvatus)、乳酸乳球菌乳酸亚种(Lactococcus lactis subsp.lactis)、冷明串珠球菌(Leuconostoc gelidum)、格氏乳杆菌(Lactobacillus gasseri)、詹氏乳杆菌(Lactobacillus jensenii)和嗜淀粉乳杆菌(Lactobacillus amylohilus)。进一步采用产酸能力、食盐耐受性、亚硝酸盐耐受性、生长温度以及蛋白质和脂肪降解能力等试验测定菌株发酵性能,最终优化结果表明乳酸乳球菌乳酸亚种具备优良乳酸菌发酵剂的性能。     

The influence of douchi starter cultures on the composition of extractive components, microbiological activity, and sensory properties of fermented fish pastes

Three experiments were conducted to test the hypothesis that douchi cultures could serve as a potential starter for enhancing the quality attributes of fermented silver carp meat. In experiment 1, an active, prefermented douchi culture was incorporated into a fish paste to aid in the fish fermentation (30 d) and facilitate biochemical production of extractive flavor components (PRF). In experiment 2, a fully fermented (30 d) douchi was added to a fish paste and the mixture was fermented for 30 d (PSF). In experiment 3, a fish paste without the douchi culture was fermented for 30 d (CF). Total extracted free amino acids increased by 68.0, 68.6, and 78.8% (P < 0.05) from their initial levels to 2930, 2422, and 1573 mg/mL after 30 d of fermentation for PRF, PSF, and CF fish pastes, respectively, of which, glutamic acid, aspartic acid, alanine, lysine, and leucine were the major amino acids (>100 mg/mL). The concentrations of both formaldehyde-reactive nitrogen and ammonia nitrogen extractives increased significantly (P < 0.05) during fermentation, following the order of PRF > PSF > CF. Low amounts of biogenic amines (<25 ppm) were produced in all samples. Sensory panel evaluation showed that PRF fish pastes had desirable aroma and taste. The douchi-inoculated fermentation could be a novel technique for expanding the utilization, consumption, and the economic values of silver carp meats. Practical Application: Douchi, a fermented soybean product, is a traditional food flavoring ingredient commonly used in China, Japan, and other Asian countries. It is also used in many Chinese cuisines in the United States. On the other hand, fermented seafood made from freshwater fish such as silver carp is known to contain bioactive components believed to promote health. The findings from the present study indicated that douchi as a novel starter can be used to produce fermented silver carp fish pastes with excellent flavor and consumer acceptability. The results may be applicable to other fish species to produce similar fermentation products.     

梅香鱼发酵菌株的筛选及对品质的影响

为有效控制产品的品质,从自然发酵的梅香鱼中分离筛选适合作为发酵剂的菌株,并研究不同菌种之间的配比对发酵产品品质的影响。结果表明,筛选出的嗜盐四联球菌（Tetragenococcus halophilus）、木糖葡萄球菌（Staphylococcus xylosus）及腐生葡萄球菌（Staphylococcus saprophyticus）符合发酵要求。与单菌株发酵及未接菌的对照组相比,3 株菌制得的混合发酵剂（比例为1∶1∶1）制备的发酵鱼感官上总体可接受程度最优,其咀嚼性和弹性也最高,并且组胺和挥发性盐基氮（total volatile basic nitrogen,TVB-N）的产生得到了有效抑制。该研究可为实现定向接种、人工控制以及规模化工业生产提供参考。     

Presence of lactobacilli in the intestinal content of freshwater fish from a river and from a farm with a recirculation system

Lactobacilli are Gram-positive and catalase negative rods commonly found in lactic acid fermented foods and in the gastrointestinal tract of mammals and birds. Few studies have described lactobacilli in freshwater fish. We analysed the presence of lactobacilli in the intestines of young and adult freshwater fish inhabiting a river environment and from fish reared in an aquaculture unit with a water recirculation system. Various species of lactobacilli were present in relatively high number in the intestines of edible fresh water fish from the river, especially in the warm season but in low numbers in the cold season. Lactobacilli were scarcely found in the intestines of edible farmed fish reared in a recirculation system in warm water. Lactobacilli are reported for the first time from the intestines of wild European eel, perch, rudd, ruffe, bleak, silver bream, chub, somnul and farmed African catfish. The two first fishes, and the last one are highly valuable species for fisheries and aquaculture. Additionally, improved methods for storage and bacteriological analysis of fish intestinal content are described. The natural presence of lactic acid bacteria in fish may be of great interest in producing fermented fish products worldwide.     

云南淡水鱼中单增李斯特氏菌污染状况及耐药性分析

目的了解云南淡水鱼中单增李斯特氏菌污染状况及耐药情况。方法从云南省昆明市、曲靖市、大理州、红河州4个淡水鱼主产区采集淡水鱼样品110件,参照GB 4789.30—2016《食品安全国家标准食品微生物学检验单核细胞增生李斯特氏菌检验》分离鉴定单增李斯特氏菌,并进一步开展16S rRNA序列鉴定和同源性分析,对鉴定菌株采用微量肉汤稀释法分析抗生素敏感性。结果110件样品检出2株单增李斯特氏菌(DZ-054、DZ-101),检出率为1.82%。2株单增李斯特氏菌之间相似度达到99.64%。DZ-101菌株对氨苄西林、青霉素敏感(S),DZ-054菌株对抗生素不敏感。结论云南省淡水鱼中单增李斯特菌的污染率较低,但仍有一定的安全隐患,相关部门应加强监管。     

臭鳜鱼源清酒乳杆菌的分离鉴定及其在臭鳜鱼发酵中的应用

利用含溴甲酚绿的MRS鉴别培养基,从传统臭鳜鱼中分离筛选到1 株乳酸菌,经形态学、生理生化实验以及16S rDNA序列分析鉴定为清酒乳杆菌,并命名为SMF-L5。清酒乳杆菌SMF-L5在MRS液体培养基中生长良好,14 h进入生长稳定期,活菌数最高可达7.0×109 CFU/mL,培养24 h后乳酸产量达10 g/L,最适培养条件为：培养温度30.9 ℃、初始pH 6.15、接种量1.94%。菌株SMF-L5具有耐受0.08 g/mL NaCl的能力,对大肠杆菌、金黄色葡萄球菌等菌株具有明显的抑制作用。接种清酒乳杆菌SMF-L5发酵的臭鳜鱼,比自然发酵臭鳜鱼具有更好的色泽、质构和风味品质。研究结果表明,臭鳜鱼源清酒乳杆菌SMF-L5可作为优良的乳酸菌发酵剂,在臭鳜鱼工业化生产中具有应用潜力。     

中国内陆6省(自治区)淡水鱼养殖、销售和餐饮环节常见嗜盐性弧菌污染调查

目的了解内陆6省(自治区)淡水鱼养殖、销售、餐饮环节鱼、水体、水底沉积物等样品中副溶血性弧菌、溶藻弧菌、创伤弧菌的污染状况,明确淡水鱼养殖、销售、餐饮各环节中3种嗜盐性弧菌的分布。方法在广西、河南、湖北、山东、四川、浙江6个省(自治区)的内陆地市设置采样点,采集养殖场、销售场所、餐馆的鱼样及存养鱼的对应水样。采用传统的分离培养技术检测3种嗜盐性弧菌,并进行生化鉴定。结果养殖环节水体和淡水鱼中副溶血性弧菌的检出率分别为1.89%(5/264)、5.81%(14/241),溶藻弧菌检出率分别为1.89%(5/264)、4.98%(12/241),创伤弧菌检出率分别为1.14%(3/264)、0.83%(2/241);3种嗜盐性弧菌的污染率在流通环节增高,存养水体和淡水鱼的检出率分别为副溶血性弧菌18.47%(29/157)、14.93%(40/268),溶藻弧菌10.83%(17/157)、9.70%(26/268),创伤弧菌3.18%(5/157)、3.36%(9/268);餐饮环节淡水鱼中嗜盐性弧菌的检出率与流通环节基本一致。结论传统意义上海水中常见的副溶血性弧菌、创伤弧菌、溶藻弧菌3种嗜盐性弧菌已经在内陆地区的淡水养殖、流通和餐饮各个环节中检出,尤其是淡水养殖环境中嗜盐性弧菌的出现可能对整个食物链产生的影响值得关注。     

Preliminary examinations on the enterotoxigenicity of isolates of Klebsiella pneumoniae from seafoods.

One hundred and eighty-five seafood samples, consisting of 96 freshwater fish, 37 marine fish, 13 freshwater prawn, 13 marine prawn and 26 molluscs were screened for presence of Klebsiella. Out of these, 12 isolates of Klebsiella were identified, Four K. pneumoniae var. ozaenae were isolated from marine fish samples and eight K. pneumoniae var. pneumoniae, six from freshwater fish and two from freshwater prawns. All 12 isolates were tested for enterotoxigenicity by the vasopermeability factor test in rabbits, the mouse foot pad test, the latex agglutination test and the coagglutination test. One isolate of K. pneumoniae var pneumoniae, isolated from fresh water prawn was found enterotoxigenic.     

传统发酵鱼制品品质及安全性研究进展

发酵鱼制品不仅提高了淡水鱼的利用率,而且赋予鱼肉较高的营养价值和良好的风味。但传统发酵鱼制品制作工艺仅依靠工人经验进行操作,导致产品质量不稳定,安全性得不到保障。本文介绍传统发酵鱼制品工艺制作过程,分析影响发酵鱼制品品质的主要因素、产品品质变化过程和安全性,旨在为发酵鱼制品的加工工艺提供新思路,以期为发酵鱼制品产业化、标准化生产提供指导。     

Selection of Autochthonous Strains as Starter Cultures for Fermented Fish Products

This paper was the 1st research focusing on the design of a halophilic lactic starter for the production of fermented fish products using a quantitative approach, based on the evaluation of the growth index and acidification score, as well as on the use of a multivariate approach to select the most promising strains. Fifty‐nine strains were randomly selected from salted fish and phenotypically characterized through Gram staining, catalase activity, glucose metabolism, H₂S and indole production, nitrate reduction, citrate utilization, and hydrolysis of arginine, esculin, casein, gelatin, starch, Tween 80, and urea. Then the Gram positive isolates (44 out of 59) were studied for their growth at different temperatures (10, 25, 40, and 55 °C), salt (0%, 20%, and 30%), pHs (4.5 and 9.5), and acidification score in lab medium. Data were modelled through growth index and used as input to run a preliminary cluster analysis and a principal component analysis. Three promising strains were selected, identified as members of the genus Pediococcus and used for the validation at laboratory level through the assessment of their performances in the production of a fermented fish sauce. The results were really promising as their use not only reduced the fermentation time (2 d) but also improved the microbiological quality of the final product. This paper represents a 1st report on the use of a simple step‐by‐step methodology to select promising halophilic strains for the optimization of a starter for fish‐fermented products.