Characterization of macrophage-activating lactic acid bacteria isolated from Mukeunji

Food Science and Biotechnology - Lactic acid bacteria (LAB) were reported to comprise the majority of the bacterial population in mukeunji, a long-fermented kimchi. This current study investigated...     

A survey of the lactic acid bacteria isolated from Serbian artisanal dairy product kajmak

Kajmak is an artisanal Serbian dairy product made by fermentation of milk fat. Overall, 374 bacterial isolates were collected from six kajmak samples of different ages produced in the households located in distinct regions of Serbia. In order to identify lactic acid bacteria present in chosen samples of kajmak, total 349 Gram-positive and catalase-negative isolates were analyzed. The recognition of isolates was performed by phenotypic characterization followed by molecular identification using (GTG)(5)-PCR and sequence analysis of 16S rRNA gene. Leuconostoc mesenteroides and Enterococcus faecium were the most frequently isolated species from kajmak samples. In contrast, leuconostocs and enterococci were found in BGMK3 and BGMK1 kajmak respectively, only after using enrichment technique for isolation suggesting they are present in low numbers in these kajmaks. Lactococcus lactis, Lactococcus raffinolactis and Lactococcus garvieae were also found in those samples but in lower proportion. Results showed that Lactobacillus plantarum, Lb. paracasei and Lb. kefiri were the most frequently isolated Lactobacillus species in analyzed kajmaks.     

Identification of predominant lactic acid bacteria isolated from traditionally fermented vegetable products of the Eastern Himalayas

Gundruk, sinki and khalpi are lactic-fermented vegetable products of Sikkim in India, and inziangsang is a fermented leafy vegetable product of Nagaland and Manipur in India. A total of 65 samples of gundruk (25), sinki (12), khalpi (25) and inziangsang (3) were analysed for microbial counts. The population of lactic acid bacteria (LAB) as well as aerobic mesophilic counts were at the level of 10(7) cfu g(-1). Yeasts were detected only in few samples of sinki and khalpi. No moulds were detected. In order to identify the predominating organisms, a total of 269 strains of LAB were isolated from gundruk, sinki, khalpi and inziangsang samples. The phenotypic characteristics of these strains were determined followed by genotyping using RAPD-PCR, repetitive element PCR and species-specific PCR techniques. The major representatives of the LAB involved in these fermentations were identified as Lactobacillus brevis, Lactobacillus plantarum, Pediococcus pentosaceus, Pediococcus acidilactici and Leuconostoc fallax.     

Characterization of lactic acid bacteria isolated from vacuum-packed cooked ring sausages

The bacterial populations of the surface layer and the centre of 15 spoiled vacuum-packed cooked ring sausages were characterized. About 95% of the total bacterial population in the surface layer and 55% at the centre were lactic acid bacteria. Another large bacterial group at the centre consisted of Bacillus spp. The lactic acid bacteria on the surface and at the centre were quite similar. Atypical streptobacteria, i.e. homofermentative psychrotrophic lactobacilli, were a major group of lactic acid bacteria in the surface layer of the spoiled sausages. Three main homofermentative groups could be observed on the basis of different carbohydrate patterns. Heterofermentative lactic acid bacteria belonged mainly to genus Leuconostoc. The proportion of leuconostocs in the spoiled sausages was also quite large. They could be divided into three main groups on the basis of different carbohydrate fermentation patterns. The lactic acid bacteria population of spoiled cooked ring sausages thus seemed to be heterogeneous. The strains isolated resembled strains observed by other workers in meat and meat products.     

Gas‐producing and spoilage potential of Enterobacteriaceae and lactic acid bacteria isolated from chilled vacuum‐packaged beef

This study aimed to enumerate and identify lactic acid bacteria and Enterobacteriaceae from spoiled and nonspoiled chilled vacuum‐packaged beef and determine their potential to cause blown pack spoilage. These microbial groups were also enumerated in nonspoiled samples and detected in abattoir samples. The potential of isolates to cause ‘blown pack’ spoilage of vacuum‐packaged beef stored at chilled temperature (4 °C) and abuse temperature (15 °C) was investigated. Populations of lactic acid bacteria in exudate of spoiled and nonspoiled samples were not significantly different (P > 0.05), whereas the number of lactic acid bacteria on the surface was significantly higher (P < 0.05) in spoiled samples as compared to nonspoiled samples. The population of Enterobacteriaceae species in exudate and on the surface of samples were significantly higher (P < 0.05) in spoiled packs in comparison with nonspoiled packs. Results of the deterioration potential showed that ‘blown pack’ spoilage was noticeable after 7 days at 15 °C and after 6 weeks at 4 °C for samples inoculated with Hafnia alvei.     

Phenotypic and technological diversity of lactic acid bacteria and staphylococci isolated from traditionally fermented sausages in southern Greece

The physicochemical and microbiological characteristics of spontaneously fermented sausages made by two medium-sized enterprises (MSE) located in southern Greece have been studied. A total of 300 lactic acid bacteria and 300 staphylococcal strains have been isolated and identified by their physiological characteristics. Lactobacillus plantarum strains were found to dominate the lactic acid bacteria microbiota in most of the cases with L. sakei strains prevailing in some of them and L. rhamnosus strains occasionally accompanying the dominant lactic acid bacteria microbiota. On the other hand, S. saprophyticus strains were found to dominate the staphylococcal microbiota in all spontaneously fermented sausages with of S. simulans, S. xylosus, S. gallinarum and S. cohnii cohnii strains being sporadically present. Following the identification, an evaluation of their technological properties, namely proteolytic and lipolytic capacities as well as production of biogenic amines and antimicrobial compounds, took place. None of the lactic acid bacteria and staphylococci was found to possess lipolytic activity whereas a total of 6 lactic acid bacteria and 51 staphylococci strains were found to be able to hydrolyse either the sarcoplasic, myofibrillar or both protein fractions. Furthermore, only one L. sakei strain and 185 staphylococci strains were found to possess decarboxylase activity against lysine, tyrosine, ornithine or histidine. Finally none of the staphylococcal microbiota and 3 lactic acid bacteria strains were found to be able to produce antimicrobial compounds of proteinaceous nature against Listeria monocytogenes.     

乳酸菌拮抗食源性致病菌研究进展

食品中致病菌是引起人类食源性疾病等食品安全问题主要因素之一。乳酸菌通过生态位竞争、形成酸性环境和产生各种代谢产物对食品中致病菌具有较强的拮抗作用。本文对乳酸菌生物保护剂的拮抗机制、作用的特点和优势,乳酸菌对各类食源性致病菌的拮抗作用研究以及在乳制品、肉制品、海产品和果蔬制品中的应用进行综述,为充分挖掘乳酸菌资源,利用乳酸菌生物保护剂控制食品中致病菌提供借鉴与参考。      

Characterization of anti-listerial lactic acid bacteria isolated from Thai fermented fish products

Thai fermented fish products were screened for lactic acid bacteria capable of inhibitingListeriasp. (Listeria innocua). Of 4150 assumed lactic acid bacteria colonies from MRS agar plates that were screened by an agar-overlay method 58 (1.4%) were positive. Forty four of these strains were further characterized and 43 strains were inhibitory againstListeria monocytogenes. The strains were inhibitory to other Gram-positive (lactic acid) bacteria probably because of production of bacteriocins. All 44 strains inhibited bothVibrio choleraeandVibrio parahaemolyticusand 37 were inhibitory to a mesophilic fish spoilage bacterium (anAeromonassp.). Inhibition of Gram-negative bacteria was attributed to production of lactic acid. Most strains were identified asLactobacillusspp., and all grew well at ambient temperatures (25–37°C) and tolerated up to 6.5% NaCl. Glucose was fermented rapidly in laboratory media whereas pH decreased only very slowly in fish juice supplemented with 4% glucose and 3.5% NaCl or in a rice–fish mixture. Only four of 44 isolates could degrade and ferment complex carbohydrates such as rice, potatoes and maize starch. This indicates that other types of bacteria may be responsible for the rapid spontaneous fermentation of the products or that other yet-unknown factors ensure rapid fermentation. Overall anti-listerial lactic acid bacteria do occur in fermented fish products and the antibacterial activity against pathogenic bacteria indicates that they may be important in product safety.     

Amino acid profiles of lactic acid bacteria, isolated from kefir grains and kefir starter made from them

The characteristics of cell growth, lactic acid production, amino acid release and consumption by single-strain cultures of lactic acid bacteria (isolated from kefir grains), and by a multiple-strain kefir starter prepared from them, were studied. The change in the levels of free amino acids was followed throughout the kefir process: single-strain kefir bacteria and the kefir starter (Lactococcus lactis C15-1%+Lactobacillus helveticus MP12-3%+(Streptococcus thermophilus T15+Lactobacillus bulgaricus HP1 = 1:1)-3%) were cultivated in pasteurized (92 degrees C for 20 min) cow's milk (3% fat content) at 28 degrees C for 5 h (the kefir starter reached pH 4.7) and subsequently grown at 20 degrees C for 16 h; storage was at 4 degrees C for 168 h. The strain L. helveticus MP12 was unrivaled with respect to free amino acid production (53.38 mg (100 g)(-1)) and cell growth (17.8 x 10(8) CFU ml(-1)); however, it manifested the lowest acidification activity. L. bulgaricus HP1 released approximately 3.7 times less amino acids, nearly 5 times lower cell growth, and produced about 1.2 times more lactic acid. S. thermophilus T15 demonstrated dramatically complex amino acid necessities for growth and metabolism. With L. lactis C15, the highest levels of growth and lactic acid synthesis were recorded (18.3 x 10(8) CFU ml(-1) and 7.8 g l(-1) lactic acid at the 21st hour), and as for free amino acid production, it approximated L. bulgaricus HP1 (17.03 mg (100 g)(-1) maximum concentration). In the L. lactis C15 culture, the amino acids were used more actively throughout the first exponential growth phase (by the 10th hour) than during the second growth phase. The unique properties of the L. helveticus MP12 strain to produce amino acids were employed to create a symbiotic bioconsortium kefir culture, which, under conditions of kefir formation, enhanced lactic acid production and shortened the time required to reach pH 4.7; intensified cell growth activity, resulting in a respective 90- and 60-fold increase in the concentration of lactobacilli and cocci in the mixed culture compared to individual cultures; and accumulated free amino acids in the final kefir with higher total concentrations (56.88 mg (100 g)(-1)) and an individual concentration of essential amino acids (1.5 times) greater than that of yogurt.     

Phenotypic identification and technological properties of lactic acid bacteria isolated from traditionally processed fish products of the Eastern Himalayas

Sukako maacha, gnuchi, sidra and sukuti are traditional smoked and sun-dried fish products of the Eastern Himalayan regions of Nepal and India. A total of 40 samples of sukako maacha (14), gnuchi (6), sidra (10) and sukuti (10) were collected and were analysed for microbial load. Population of lactic acid bacteria (LAB) as well as aerobic mesophilic counts ranged from 4.7-8.3 to 5.1-8.5 log cfu g(-1), respectively. A total of 189 strains of LAB were isolated from sukako maacha, gnuchi, sidra and sukuti samples, out of which 171 strains were cocci and 15 strains, were heterofermentative lactobacilli. LAB were identified on the basis of phenotypic characters including API system as Lactococcus lactis subsp. cremoris, Lactococcus lactis subsp. lactis, Lactococcus plantarum, Leuconostoc mesenteroides, Enterococcus faecium, Enterococcus faecalis, Pediococcus pentosaceus and Weissella confusa. LAB strains produced a wide spectrum of enzymes. Some strains of LAB showed antagonistic properties against pathogenic strains. None of the strains produced biogenic amines in the method applied. This paper is the first report on the microbial composition, mostly lactic acid bacteria, of traditionally processed fish products of Eastern Himalayas.     

Identification of cultivable lactic acid bacteria isolated from Algerian raw goat's milk and evaluation of their technological properties

One hundred and fifty-eight strains of lactic acid bacteria isolated from Algerian raw goat's milk were identified and technologically characterized. Five genera were found: Lactobacillus (50.63%), Lactococcus (25.94%), Streptococcus (14.56%), Leuconostoc (7.59%) and Pediococcus (1.26%). The predominant species were Lactococcus lactis (32 strains), Streptococcus thermophilus (23 strains), Lactobacillus bulgaricus (19 strains), Lb. helveticus (16 strains) and Lb. plantarum (14 strains).Approximately 39% of the lactic acid bacteria isolated produced more than 0.6% lactic acid (w/v) after 18 h of incubation, and belonged to the Lactococcus and Lactobacillus genera. The highest proteolytic activity was approximately 3 mg tyrosine l⁻¹ for mesophilic strains and nearly 5 mg tyrosine l⁻¹ for thermophilic lactobacilli after 72 h. High aromatic activity (more than 0.8 mg diacetyl l⁻¹ after 16 h) was detected in 14% of the strains.Nine strains were used to make dairy products (a yoghurt-like product and Edam-type cheese) on a pilot scale in the laboratory. The best-liked organoleptic characteristics were noted in a yoghurt produced with a mixed culture made up of S. thermophilus (strain 16TMC+) and Lb. helveticus (strain 20TMC) and in a cheese made with a starter composed of Lc. lactis subsp. lactis (strain 10MCM) and L. lactis subsp. lactis (V.P. +) (strain 19MCM).     

Screening of lactic acid bacteria isolated from fermented table olives with probiotic potential

The aim of this work was to study the potential probiotic properties of lactobacilli associated with table olives. From a total of 111 isolates from spontaneously fermented green olive brines, 109 were identified at species level by multiplex PCR amplifications of the recA gene. One hundred and seven of these were identified as Lactobacillus pentosus, one as Lactobacillus plantarum, and another as Lactobacillus paraplantarum. Repetitive bacterial DNA element fingerprinting (rep-PCR) with GTG₅ primer revealed a higher variability within the L. pentosus isolates and nine different clusters were obtained. Most of them showed high autoaggregation ability, low hydrophobicity properties, and lower survival to gastric than to pancreatic digestion; however, no isolate showed bacteriocin, haemolytic or bile salt hydrolase activities. A multivariate analysis based on results from phenotypic tests led to the segregation of some L. pentosus isolates with promising potential probiotic characteristics, which are even better than probiotic reference strains. Due to the autochthonous origin of the strains, their use as starter cultures may contribute to improving natural fermentation and the nutritional characteristics of table olives.     

黑龙江传统发酵豆酱中乳酸菌的分离鉴定

为了探明和保护黑龙江传统发酵豆酱中丰富的乳酸菌资源,以采自黑龙江农家12份传统发酵豆酱样品为试材,从中选择性培养分离出豆酱中的优势乳酸菌疑似菌株,再利用16S rDNA序列分析方法,对其进行鉴定并保藏。结果表明,从黑龙江3个地区采集到的12份样品中共分离出了24株乳酸菌,通过16S rDNA序列分析后共鉴定出5个乳酸菌种,分别是嗜盐四联球菌(T.halopHilus)10株,占总数的41.67%;植物乳杆菌(L.plantarum)6株,占总数的25%;清酒乳杆菌(L.sakei)4株,占总数的16.67%;发酵乳杆菌(L.fermentum)2株,占总数8.33%;短乳杆菌(L.brevi)2株,占总数8.33%。其中,在这3个地区采集的样品中均分离到了嗜盐四联球菌、植物乳杆菌和清酒乳杆菌,嗜盐四联球菌在其中的9份样品中分离到,占总样品数的75%。植物乳杆菌在其中的6份样品中分离到,占总样品数的50%。可以初步推断嗜盐四联球菌和植物乳杆菌是黑龙江传统发酵豆酱中优势乳酸菌菌群。     

传统发酵蔬菜中拮抗温和气单胞菌的乳酸菌的筛选与鉴定

目的:从传统发酵蔬菜中筛选对温和气单胞菌具有拮抗作用的乳酸菌。方法:采用双层琼脂扩散法筛选,通过生理生化反应和16S rRNA序列分析进行鉴定,研究蛋白酶、p H和温度等因素对乳酸菌无细胞上清液（CFS）抑菌活性的影响,采用扫描电镜分析乳酸菌无细胞上清液对温和气单胞菌细胞结构完整性的影响。结果:从腌渍酸黄瓜中筛选出对温和气单胞菌（10⁶CFU/m L）具有较强抑制活性的菌株LZH2-5,经鉴定为植物乳杆菌（Lactobacillus plantarum）。LZH2-5无细胞上清液经胰蛋白酶、木瓜蛋白酶、中性蛋白酶、碱性蛋白酶和胃蛋白酶处理后其抑菌活性分别丧失29.38%、23.93%、22.30%、19.47%和14.59%;经40、60、80℃处理30 min后其拮抗活性基本不变,在100℃和121℃处理30 min后活性分别下降了16.66%和25.40%,在p H3.0⁴.0范围内保持其抑菌活性,粗提物的最小抑菌浓度12.0 mg/m L。扫描电镜表明经LZH2-5 CFS处理使温和气单胞菌的细胞结构破坏并溶解。结论:从腌渍酸黄瓜中筛选的植物乳杆菌LZH2-5对温和气单胞菌具有较强抑制作用,初步分析抑菌活性物质为细菌素类,可作为水产品养殖过程中控制温和气单胞菌的拮抗制剂出发菌株。      

Characterization of lactic acid bacteria isolated from a Greek dry-fermented sausage in respect of their technological and probiotic properties

A total of 147 lactic acid bacteria was isolated from two types of naturally fermented dry sausages at four different stages of the ripening process studied in order to select the most suitable strains according to their technological characteristics including probiotic properties and antimicrobial activity against food-borne pathogens. Identification of the isolates revealed that 90% were lactobacilli, 4% enterococci, 3% Pediococcus sp. and sporadic isolates of Weissella viridescens, Leuconostoc pseudomesenteroides, and Leuconostoc sp. The isolated strains of Lactobacillus sakei (49 isolates), Lactobacillus curvatus (24 isolates) and Lactobacillus plantarum (7 isolates) were further characterized. All strains could grow at 15?°C, whereas the majority of the strains was able to grow in the presence of 6.5% NaCl and on acetate agar. The enzymatic potential of the strains was evaluated using the API ZYM system. During in vitro investigations all strains exhibited high leucine and valine aminopeptidase activities and moderate acid phosphatase and phosphohydrolase activities. Some strains showed very weak lipolytic activity. The enzyme profiling is an important factor for selection of strains as starter cultures. A large majority of the strains tolerated 0.1% bile salts whereas 58% of Lactobacillus curvatus strains and all Lactobacillus plantarum strains were resistant to 0.3% bile salts. All Lactobacillus sakei strains and the majority of Lactobacillus curvatus and Lactobacillus plantarum strains exhibited an anti-listerial activity against three Listeria monocytogenes strains. A percentage of 75, 50 and 29% of Lactobacillus sakei, L. curvatus and L. plantarum strains, respectively, could inhibit two Staphylococcus aureus strains. The contribution of the selected strains to a possible inhibition of Listeria monocytogenes and S. aureus in situ on fermented meats would be of considerable interest to enhance the hygienic quality of these products.     

In vitro evaluation of probiotic lactic acid bacteria isolated from dairy and non-dairy environments

Four strains and 2 strains of lactic acid bacteria (LAB) were isolated from the commercial yogurt and kimchi products in Korea, respectively. Based on the 16S rRNA sequencing data, strain A from a drink-type yogurt manufactured by dairy company S, was a Gram-positive, rod-shaped Lactobacillus helveticus, and both strain B (company N) and D (company H) were identified as L. casei ssp. casei, and strain C (company L) as L. paracasei. None of yogurt strain B and D was recovered from the samples exposed to the simulated gastric juice, pH 2.0 for 1.5 hr. Of the 6 isolates tested, strain YS93 from kimchi was the most resistant to acidic condition using the simulated gastric juice, pH 2.0. Moreover, it was shown that 2 kimchi isolates and yogurt strain D produced antibacterial substances, probably bacteriocin-like peptide, which was inhibitory against Staphylococcus aureus as an indicator. In an adhesion assay using a Caco-2 cell, the adherence activity of kimchi strains YS29 and YS93 was significantly higher than those of 4 yogurt starter strains tested.     

高产EPS乳酸菌株的筛选及在农家干酪中的应用

从16株乳酸菌中筛选出三株高产胞外多糖菌种,分别为嗜热链球菌S7和保加利亚乳杆菌L6及LTM,应用苯酚—硫酸法测定其胞外多糖的产量分别为56.25、51.53、51.48mg/L,产量相对较高且稳定。利用菌株S7、L6和LTM制作农家干酪的研究表明,以感官评定值为主指标,结合农家干酪的含水量、蛋白质含量和硬度进行分析,确定当稀奶油添加量为5.2%、72℃热处理15min、氯化钙添加量为0.01%时,生产的农家干酪品质最佳。      

Characterization of lactic acid bacteria isolated from Bukuljac, a homemade goat's milk cheese

The Bukuljac cheese is traditionally homemade cheese, produced from heat-treated goat's milk without the addition of any bacterial starter culture. The presence of lactic acid bacteria (LAB) in Bukuljac cheese has been analyzed by using a polyphasic approach including microbiological and molecular methods such as rep-PCR with (GTG)5 primer. Lactobacillus paracasei subsp. paracasei represents a dominant strain in the microflora of analyzed cheese. Out of 55 Gram-positive and catalase-negative isolates, 48 belonged to L. paracasei subsp. paracasei species. Besides lactobacilli, five Lactococcus lactis subsp. lactis and two Enterococcus faecalis were found. Results of PCR-denaturing gradient gel electrophoresis (DGGE) of DNA extracted directly from the fresh cheese revealed the presence of Leuconostoc mesenteroides. Only lactobacilli showed a high proteolytic activity and hydrolyzed alpha(s1)- and beta-caseins. They are also producers of diacetyl. In addition, 34 out of 55 isolates, all determined as lactobacilli, showed the ability of auto-aggregation. Among 55 isolates, 50 also exhibited antimicrobial activity.     

In vitro probiotic evaluation of potential antioxidant lactic acid bacteria isolated from idli batter fermented with Piper betle leaves

Lactic acid bacteria isolated from food sources can be introduced as probiotics because of their health‐promoting and nonpathogenic characteristics. Eight lactic acid bacteria from idli batter fermented with Piper betle leaves were selected to screen the antioxidant potential and biofilm‐forming ability. Five isolates exhibited good antioxidant potential and biofilm formation and showed antagonistic activity against the pathogenic biofilm‐forming Staphylococcus aureus. Further, in vitro probiotic properties of five isolates exhibited excellent acid tolerance, bile tolerance, simulated gastrointestinal juice tolerance, auto‐aggregation and co‐aggregation and hydrophobicity and showed resistance to antibiotics such as ciprofloxacin, nalidixic acid, norfloxacin and colistin. Isolates also showed positive bile salt hydrolase activity, cholesterol assimilation, β‐galactosidase production and mucin binding ability. The isolate KJBB10 exhibited good adhesion and pathogen exclusion percentage with human colon cancer cells (HCT‐15 and HT‐29). Hence, the isolates KJBB10, KJBB56 and KJBC06 can be used as beneficial probiotic starter cultures for the formulation of functional foods.     

腐乳中乳酸菌的分离与鉴定

文章主要对青方、白方和红方腐乳中的乳酸菌进行了分离鉴定。从青方和白方腐乳中分别分离到乳酸菌7株和5株,红方中没有分离到乳酸菌。鉴定结果表明,白方中4株为鼠李糖乳杆菌,1株清酒乳杆菌;青方中有1株为植物乳杆菌,1株干酪乳杆菌,3株鼠李糖乳杆菌,2株清酒乳杆菌。最后,对不同菌株的耐盐性和耐酒精性进行了测试。