Evaluation of stored lamb bio-preserved using a three-strain cocktail of Lactobacillus sakei

Commercially prepared lamb was stored at −1.5 °C after inoculation with a combination of three Lactobacillus sakei strains previously shown to inhibit spoilage and pathogenic bacteria of importance to the meat industry. Between 6 and 14 weeks storage samples were evaluated for growth of inoculated strains, production of fermentation end-products and sensory acceptance of the cooked product. All three L. sakei strains flourished during storage, formed consistently dominant populations and were associated with lower surface pH and increased levels of lactic and acetic acids. Inoculated samples were determined to be as equally acceptable for smell, acidity, rancidity and overall liking as un-inoculated controls.     

The interaction of the non-bacteriocinogenic Lactobacillus sakei 10A and lactocin S producing Lactobacillus sakei 148 towards Listeria monocytogenes on a model cooked ham

Two lactic acid bacteria, Lactobacillus sakei subsp. carnosus (10A) and lactocin S producing Lactobacillus sakei 148 (LS5), were examined for their usefulness as protective culture in the biopreservation of cooked meat products. Co-culture experiments on a model cooked ham (MCH) between 10A or LS5 and a cocktail of three Listeria monocytogenes strains were performed to examine the influence of inoculum level (10(5) vs. 10(6)cfu/g), storage temperature (4 vs. 7 degrees C) and packaging type (vacuum-packaging vs. modified atmosphere-packaging). At 7 degrees C, applying Lactobacillus sakei 10A at 10(6) cfu/g limited the growth of Listeria monocytogenes to <1 log(10) cfu/g during 27 days, whilst an application level of 10(5) cfu/g failed to prevent growth to unacceptable levels. Lactobacillus sakei LS5 did not demonstrate an antagonistic effect towards Listeria monocytogenes. Lowering the temperature to 4 degrees C or switching from vacuum-packaging to modified atmosphere packaging (MAP) did not influence the ability of strain 10A to grow on the MCH, as its dominance did not change. A combination of strain 10A and 4 degrees C or a MAP containing 50% CO(2) completely inhibited the growth of Listeria monocytogenes. Sensory assessments and pH measurements confirmed that 10A, even when present at a high level for prolonged storage times, did not acidify the cooked ham to a point of sensory rejection.     

Isolation and characterization of a new dimeric esterase from Lactobacillus sakei 23K

The M_w of a Lactobacillus sakei intracellular esterase, determined by gel filtration, was compared to those obtained from SDS-PAGE or MALDI-TOF, pointing to a dimeric structure. Its N-terminal sequence and peptide mass fingerprint suggest that it is the putative LSA044 protein from L. sakei 23K genome.     

Technological characterization of a bacteriocin-producing Lactobacillus sakei and its use in fermented sausages production

The aim of this paper was the technological characterization of a Lactobacillus sakei strain, able to produce the bacteriocin sakacin P, that was originally isolated from naturally fermented sausages. Experiments were conducted in situ, using MRS-based medium, and in situ, when the strain was inoculated as starter culture in real sausage fermentation. The results obtained underlined that the strain was able to grow in conditions that are commonly used in the production line, and only lactose and high concentrations of NaCl (5% w/v) reduced the capability for bacteriocin production. When inoculated in sausages, the strain showed a good performance, being able to colonize rapidly the ecosystem. A high number of isolates, capable of producing sakacin P, were already isolated after the third day of fermentation, and persisted throughout the course of the fermentation. The inoculated strain also affected other microbial colonization trends; in fact the total bacterial count and fecal enterococci showed a rapid decrease at the end of the fermentation. Moreover, during sensory evaluation, the final sausage product received high scores for the parameters of tenderness and juiciness, with medium acidity and low rancidity. Lastly, the panelists preferred the sausages produced with the L. sakei characterized in this study when compared to a fermented sausage produced with a commercial starter.     

清酒乳杆菌对发酵风干肠品质的影响

该研究采用清酒乳杆菌（Lactobacillus sakei，L. sakei）作为发酵菌株制备发酵风干肠，研究其对发酵风干肠品质的影响，以自然发酵产品作为对照组，在发酵成熟过程中的第0、3、6、9天测定产品的菌落总数、乳酸菌数、pH值、水分含量、水分分布、嫩度、色差、过氧化物值（Peroxide Value，POV）、硫代巴比妥酸值（Thiobarbituric Acid Value，TBARS）、脂肪酸含量和感官评分等指标，研究发酵菌株对风干肠品质的影响。试验表明，L. sakei接种组的乳酸菌数和菌落总数显著高于对照组（P<0.05），L*值、a*和b*显著低于对照组（P<0.05）；L. sakei接种组在发酵终点时，水分含量降至32.15%，pH值降至4.41，保证了发酵风干肠的安全性；POV值和TBARS值显著低于对照组（P<0.05），表明L. sakei具有较好的抑制脂肪氧化作用；L. sakei接种组的总体可接受性更高，发酵后期酯类物质种类和含量均高于对照组。综上，L. sakei的添加改善了产品的品质和风味。     

Metabolism of amino acids, dipeptides and tetrapeptides by Lactobacillus sakei

The microbial degradation of proteins, peptides and amino acids generates volatiles involved in the typical flavor of dry fermented sausage. The ability of three Lactobacillus sakei strains to form aroma compounds was investigated. Whole resting cells were fermented in phosphate buffer with equimolar amounts of substrates consisting of dipeptides, tetrapeptides and free amino acids, respectively.Dipeptides disappeared quickly from the solutions whereas tetrapeptides were only partially degraded. In both approaches the concentration of free amino acids increased in the reaction mixture but did not reach the equimolar amount of the initial substrates. When free amino acids were fed to the bacteria their levels decreased only slightly. Although peptides were more rapidly degraded and/or transported into the cells, free amino acids produced higher amounts of volatiles.It is suggested, that after transport into the cell peptides are only partially hydrolyzed to their amino acids, while the rest is metabolized via alternative metabolic pathways. The three L. sakei strains differed to some extend in their ability to metabolize the substrates to volatile compounds. In a few cases this was due to the position of the amino acids within the peptides. Compared to other starter cultures used for the production of dry fermented sausages, the metabolic impact of the L. sakei strains on the formation of volatiles was very low.     

清酒乳杆菌对新疆酸驼乳发酵特性的影响

关于新疆酸驼乳研究已经表明,清酒乳杆菌存在于自然发酵酸驼乳中,但清酒乳杆菌的存在是否对酸驼乳发酵产生正面影响还未清楚。为此,本实验首先建立一种基于特有菌落形态与katA基因相结合的清酒乳杆菌检测方法,从而跟踪驼乳的发酵过程,用不含清酒乳杆菌酸驼乳和含有清酒乳杆菌的酸驼乳作为自然发酵剂分别接种新鲜消毒驼乳,结果表明,接种含有清酒乳杆菌的酸驼乳的新鲜驼乳其黏度随着时间延长而逐渐下降;相反接种不含清酒乳杆菌酸驼乳的新鲜消毒驼乳,其黏度随着时间延长而增高。进一步用分离自酸驼乳的清酒乳杆菌与6株其他乳酸菌分别接种新鲜消毒驼乳,结果表明:接种清酒乳杆菌的驼乳黏度均有下降;相反接种其他乳酸菌的驼乳黏度均有不同程度的增加。这一结果初步揭示了在低黏、奶香型传统驼乳发酵中清酒乳杆菌起重要作用。     

添加肉类蛋白对米酒乳杆菌生境适应相关基因表达的影响

米酒乳杆菌是常见的异型发酵乳酸菌,它常常用作肉类工业发酵剂。为了了解其在肉类环境中的生存状况,本文用定量PCR对米酒乳杆菌La22在添加肌浆蛋白和肌原纤维蛋白的培养基中生长时生境适应相关基因表达量进行分析。结果显示,在两种培养基中生长时,p H值均随时间的延长而下降,在添加肌浆蛋白的培养基略高于添加肌原纤维蛋白培养基。与对照相比,在添加肌浆蛋白培养基中的arc A、arc B、arc C、arc T以及添加肌原纤维蛋白培养基中的arc R基因表达量明显上调,其余几个arc基因也均上调,但不明显;压力相关基因lsa00513、usp4以及usp2在肌浆蛋白培养基中均明显上调;有6个ABC转运子基因在添加两种肌肉蛋白的培养基中表达量明显上调。结果表明米酒乳杆菌能通过能量转换与基因表达调节来适应含肉类蛋白的培养环境。     

In vitro evaluation of the probiotic potential of bacteriocin producer Lactobacillus sakei 1

Lactobacillus sakei 1 is a food isolate that produces a heat-stable antimicrobial peptide (sakacin 1, a class IIa bacteriocin) inhibitory to the opportunistic pathogen Listeria monocytogenes. Bacterial isolates with antimicrobial activity may be useful for food biopreservation and also for developing probiotics. To evaluate the probiotic potential of L. sakei 1, it was tested for (i) in vitro gastric resistance (with synthetic gastric juice adjusted to pH 2.0, 2.5, or 3.0); (ii) survival and bacteriocin production in the presence of bile salts and commercial prebiotics (inulin and oligofructose); (iii) adhesion to Caco-2 cells; and (iv) effect on the adhesion of L. monocytogenes to Caco-2 cells and invasion of these cells by the organism. The results showed that L. sakei 1 survival in gastric environment varied according to pH, with the maximum survival achieved at pH 3.0, despite a 4-log reduction of the population after 3 h. Regarding the bile salt tolerance and influence of prebiotics, it was observed that L. sakei 1 survival rates were similar (P > 0.05) for all de Man Rogosa Sharpe (MRS) broth formulations when tests were done after 4 h of incubation. However, after incubation for 24 h, the survival of L. sakei 1 in MRS broth was reduced by 1.8 log (P < 0.001), when glucose was replaced by either inulin or oligofructose (without Oxgall). L. sakei 1 was unable to deconjugate bile salts, and there was a significant decrease (1.4 log) of the L. sakei 1 population in regular MRS broth plus Oxgall (P < 0.05). In spite of this, tolerance levels of L. sakei 1 to bile salts were similar in regular MRS broth and in MRS broth with oligofructose. Lower bacteriocin production was observed in MRS broth when inulin (3,200 AU/ml) or oligofructose (2,400 AU/ml) was used instead of glucose (6,400 AU/ml). L. sakei 1 adhered to Caco-2 cells, and its cell-free pH-neutralized supernatant containing sakacin 1 led to a significant reduction of in vitro listerial invasion of human intestinal Caco-2 cells.     

Characterization of new bacteriocinogenic lactic acid bacteria isolated using a medium designed to simulate inhibition of Listeria by Lactobacillus sakei 2512 on meat

Bacteriocinogenic bacteria have been proposed to protect food products from Listeria contamination as bioprotective cultures. Lactobacillus sakei 2512 was demonstrated to inhibit the growth of Listeria on sliced cooked ham by challenge test. A liquid medium simulating ham, BHI5L200, was designed in order to select bioprotective strains for meat protection. Two strains were selected, from the 201 lactic acid bacteria screened, that produced bacteriocins at pH 5.8 in BHI5L200. The first one, Leuconostoc pseudomesenteroides 2733, produced a new bacteriocin which was purified and partially characterized. The second, Lactobacillus curvatus 2711, produced sakacin X and was shown to contain sakacin T and sakacin P structural genes. Co-culture experiments in BHI5L200 demonstrated that growth of Listeria was inhibited by L. sakei 2512 as well as by L. curvatus 2711.     

清酒乳杆菌对臭鳜鱼食用品质及挥发性风味物质的影响

接种清酒乳杆菌,与自然发酵组作对比,分析菌株对臭鳜鱼发酵过程中食用品质和挥发性风味物质的影响.结果表明,接种清酒乳杆菌腌制发酵后,臭鳜鱼特征蒜瓣肉与自然发酵组同阶段相比,更规则和白亮.接种发酵组的鱼肉色泽,包括亮度和白度均优于自然发酵组,鱼肉硬度低于自然发酵组,且弹性、黏附性、胶黏性和咀嚼性均高于自然发酵组.两组发酵过...     

Feasibility of Coupling Dehydration-Impregnation by Soaking Treatment of Meat with Fermentation by Lactobacillus sakei

This study examined the feasibility of coupling dehydration-impregnation by soaking (DIS) with a subsequent lactic fermentation in the treatment of meat. A series of beef fillets were subjected to 3 different DIS treatments. The resulting DIS-treated fillets had 3 different characteristics in terms of water activity, salt, and fermentable sugars contents. Fillets treated with the DIS with the shortest immersion time (5 h) and the highest salt concentration in the DIS bath (100 g/L) were inoculated with Lactobacillus sakei. A control group was left without inoculation. After 24 h incubation at 25 °C, only inoculated fillets showed signs of lactic fermentation. At 24 h, these fillets had a d-lactic acid content of 68 μmol/g dry basis and a high population of L. sakei revealed by methods of plate count and quantitative PCR. DIS could therefore be compatible with a subsequent fermentation step by L. sakei. Practical Application: Traditional meat preservation processes often combine unit operations such as salting, smoking, fermentation, and drying. In tropical countries, high temperatures and high relative humidity, poor infrastructure, and improper slaughterhouse practices explain the need for more drastic processes (more salt, more water loss) for meat preservation. Dehydration-impregnation by soaking (DIS) could be used as a rapid pretreatment of meat, in order to counteract tropical conditions. This study validates a novel approach whereby DIS is coupled with lactic fermentation by surface inoculation with Lactobacillus sakei. With a final drying step this process could be used for the treatment of whole meat pieces.     

The control of Listeria innocua and Lactobacillus sakei in broth and meat slurry with the bacteriocinogenic strain Lactobacillus casei CRL705

The effect of the bacteriocin-producing strain, Lactobacillus casei CRL705, in the control of Listeria innocua 7 and Lactobacillus sakei CRL1424 in MRS medium and meat slurry during the storage under vacuum at chill temperatures was evaluated. L. sakei CRL 1424 isolated from vacuum-packaged contaminated raw meat was identified as the predominant indigenous lactic acid bacterial flora. Co-inoculation of MRS broth at 8°C with L. casei CRL705 caused growth inhibition of L. sakei CRL1424 and L. innocua 7 after 10 and 4 days of storage, respectively. At 4°C the complete inhibition of both strains occurred within 14 and 8 days for L. sakei CRL1424 and L. innocua 7, respectively. Bacteriocin activity in broth was observed to be maximal at 8°C reaching 2130 AU ml⁻¹ after10 days and 400 AU m1⁻¹ after 8 days of storage, while at 4°C maximal activities, 690 and 30 AU ml⁻¹ were obtained at 14 and 10 days, respectively. Addition of bacteriocinogenic strain to the meat slurry did not allow the growth of L. sakei and L. innocua, showing a bacteriostatic effect during 21 days of storage at 4°C. In addition, L. casei CRL705, as a protective culture did not change significantly the pH of the meat slurry in the assayed storage conditions. The results presented here confirm that the bacteriocinogenic strain L. casei CRL705 can be used as a useful tool to improve the microbial stability and safety in the commercial meat preservation.     

Method for reliable isolation of Lactobacillus sakei strains originating from Tunisian seafood and meat products

In Tunisia, several food products derived from meat or seafood are naturally processed, without any addition of bacterial starters. Such fermented, dried-cured, salted, or marinated products, as well as the raw meat or fish may thus provide a source to isolate the natural microflora colonizing such environments. We isolated lactic acid bacteria from a representative range of flesh-foods sold or manufactured in different parts of Tunisia, and selectively searched for Lactobacillus sakei, a lactic acid bacterium potentially useful as starter or protective culture. Eighty six (86) strains were isolated from various seafood (anchovy, sardine, sole, mullet, and octopus), or meat (pork, veal, beef, sheep, chicken, and turkey) products that were either fresh, or transformed by different traditional processes. Several methods were used in order to develop a rapid and reliable protocol for the direct identification of L. sakei. Amplified ribosomal DNA restriction analysis (ARDRA) classified the various isolates into 9 distinct groups. Search for the presence of the L. sakei specific katA gene indicated that all positive isolates were grouped in the same ARDRA group. Sequencing of 16S rDNA confirmed those isolates as L. sakei. Those 22 different L. sakei strains represent 25.6% of the total isolates, while other isolates found in the different ARDRA groups were tentatively ascribed to Lactobacillus plantarum, Lactococcus lactis/garviae, Enterococcus avium, Streptococcus parauberis, Hafnia alvei, Pediococcus pentosaceus, and Lactobacillus curvatus through 16S rDNA sequencing. A fast and reliable method to isolate and discriminate L. sakei from complex food environments is proposed.     

Co-culture experiments demonstrate the usefulness of Lactobacillus sakei 10A to prolong the shelf-life of a model cooked ham

This study investigated the usefulness of two selected lactic acid bacteria, Lactobacillus sakei subsp. carnosus (10A) and the lactocin S producing L. sakei 148 (LS5), to extend the shelf-life of cooked meat products. The interaction between these potential protective cultures and the spoilage organisms, Leuconostoc mesenteroides (LM4) and Brochothrix thermosphacta (BT1), were examined in co-culture studies on a model cooked ham product at 7 degrees C under vacuum packaged conditions. Furthermore, the influence of the glucose content of the model cooked ham on the interaction phenomena was investigated. When artificially contaminating the model cooked ham with BT1 at 10(2) cfu/g in combination with 10A at 10(5) cfu/g, the growth of BT1 was significantly slower compared to a simultaneous mono-culture experiment. In a similar experiment with LM4, LM4 reached a level of 10(7) cfu/g +/-14 days later when LM4 grew together with 10A compared to its growth in mono-culture. The lactocin S producing LS5 did not demonstrate an inhibitory action towards LM4 or BT1 and is therefore not useful as protective culture on cooked meat products. The glucose level of the model cooked ham had no influence on the observed antagonistic interactions of 10A towards LM4 or BT1, indicating that the action of the biopreservative 10A in cooked meat products is independent of the substrate glucose.     

Bioconversion of agro‐industrial by‐products to lactic acid using Lactobacillus sakei and two Pediococcus spp. strains

The aim of this study was to evaluate the bioconversion efficiency of rich in cellulose agro‐industrial by‐products such as wheat bran (WB), spent distiller's grain with solids (DGS), brewer's spent grain (BSG) and lupin (Lupinus angustifolius L.) wholemeal fraction (LF) to lactic acid (LA) using acid tolerant lactic acid bacteria (LAB) strains Lactobacillus sakei KTU05‐06, Pediococcus acidilactici KTU05‐7 and P. pentosaceus KTU05‐9. Carbohydrase preparation Depol^? 692L was used for the hydrolysis of non‐starch polysaccharides. Analysed raw materials were suitable substrates for LAB propagation and L‐lactic acid production. The lowest pH (3.6) was found in LF medium after 48 h fermentation with P. acidilactici and P. pentosaceus strains. The lowest pH (3.86) was measured in WB fermented with L. sakei, and in DGS and BSG (pH 3.8 and 3.9 respectively) fermented with P. acidilactici. The highest endoxylanase activity was excreted by the P. acidilactici and P. pentosaceus (84 and 69 XU g^?1 respectively), and the highest α‐amylase activity was of L. sakei (255.6 AU g^?1) after 24 h incubation in WB medium. The L‐lactic acid concentration of 86.11 g kg^?1 was reached after the bioconversion of hydrolysed WB in combination with 48 h fermentation by P. pentosaceus KTU05‐9 strain. LA contents between 222 and 282 mg kg^?1 was produced from lupin processing residues via fermentation using P. acidilactici and P. pentosaceus KTU05‐9 strains. The major challenge within the presented study is the viability of tested LAB in cereal waste media and effective LA production at a low pH (3.6–3.8).     

Sensory evaluation of black pigmented rice (Oryza sativa cv. Poireton) wine fortified with probiotic Lactobacillus acidophilus ATCC 4356 and Lactobacillus sakei ATCC 15521 using fuzzy logic

Rice wine was prepared in the laboratory from a black pigmented rice (cv. Poireton) using standardized methodology. The wine was fortified with the probiotic strains Lactobacillus acidophilus ATCC 4356 and Lactobacillus sakei ATCC 15521 and allowed to undergo a storage period of 28 days at 4°C. These rice wines were compared with a commercial wine used as a standard reference wine. A panel consisting of 30 judges performed the sensory evaluation. The scores obtained were based on the quality attributes of colour, taste, aroma and alcoholic strength. Analysis of the sensory scores and ranking of the different samples were performed using fuzzy analysis. The quality attributes were also ranked according to their importance in the overall acceptability. Colour and alcoholic strength were accorded the highest importance, compared with aroma and taste, as quality attributes. The rice wines were rated as ‘not satisfactory’, ‘fair’, ‘medium’, ‘good’ and ‘excellent’ on the sensory scale. Out of the wines prepared, black pigmented rice wine fortified with L. acidophilus was the most acceptable and was ranked as ‘good’, which was also at par with the commercial wine. On the other hand, black pigmented rice wine fortified with L. sakei and black pigmented rice wine fortified with L. acidophilus and L. sakei were ranked in the ‘satisfactory’ category followed by the unfortified wine ranked in the ‘fair’ category. Copyright © 2015 The Institute of Brewing & Distilling     

鲈鱼肠道LactobacillussakeiLY1-6对荧光假单胞菌抑制作用研究

目的:从海鱼肠道中筛选对荧光假单胞菌具有较强抑制作用的乳酸菌。方法:采用牛津杯琼脂扩散法筛选菌株,通过生理生化反应和16S rRNA测序进行菌株生物学鉴定,利用蛋白酶、p H和温度等因素对抑菌活性进行分析,采用扫描电镜分析乳酸菌无细胞上清液（CFS）对荧光假单胞菌细胞结构的完整性影响。结果:从鲈鱼肠道中筛选出对荧光假单胞菌具有较强抑制活性的菌株LY1-6,经生理生化和16S rRNA测序鉴定为清酒乳杆菌（Lactobacillus sakei）。菌株LY1-6 CFS经胃蛋白酶处理后其抑菌活性完全丧失,经中性蛋白酶、木瓜蛋白酶、胰蛋白酶和碱性蛋白酶处理后其抑菌活性分别丧失了38.22%、38.00%、22.48%和18.46%。在p H2.5⁵.0具有抑菌活性,并具有良好的热稳定性,初步结果表明LY1-6 CFS中抑菌活性物质可能为细菌素。扫描电镜结果表明经LY1-6 CFS处理使荧光假单胞菌细胞结构被破坏并溶解。结论:来自鲈鱼肠道的清酒乳杆菌LY1-6对荧光假单胞菌具有较强抑制作用,其抑菌活性物质为细菌素类,作用机制为破坏细胞的完整性,可作为控制水产品中荧光假单胞菌腐败生物保鲜的出发菌株。      

Evaluation of Lactobacillus rhamnosus GG and Lactobacillus acidophilus NCFM encapsulated using a novel impinging aerosol method in fruit food products

This study investigated the effect of microencapsulation on the survival of Lactobacillus rhamnosus GG and Lactobacillus acidophilus NCFM and their acidification in orange juice at 25°C for nine days and at 4°C over thirty five days of storage. Alginate micro beads (10-40 μm) containing the probiotics were produced by a novel dual aerosol method of alginate and CaCl(2) cross linking solution. Unencapsulated L. rhamnosus GG was found to have excellent survivability in orange juice at both temperatures. However unencapsulated L. acidophilus NCFM showed significant reduction in viability. Encapsulation of these two bacteria did not significantly enhance survivability but did reduce acidification at 25°C and 4°C. In agreement with this, encapsulation of L. rhamnosus GG also reduced acidification in pear and peach fruit-based foods at 25°C, however at 4°C difference in pH was insignificant between free and encapsulated cells. In conclusion, L. rhamnosus GG showed excellent survival in orange juice and microencapsulation has potential in reducing acidification and possible negative sensory effects of probiotics in orange juice and other fruit-based products.