Characterization and selection of autochthonous lactic acid bacteria isolated from traditional Iberian dry-fermented salchichón and chorizo sausages

ABSTRACT:  A total of 192 lactic acid bacteria were isolated from 2 types of naturally fermented dry sausages at 3 different stages of the ripening process in order to select the most suitable strains as starter cultures in dry-cured sausage manufacture according to their technological characteristics such as glucose fermentation, lactic and acetic acid production, and proteolytic, lipolytic, and antimicrobial activities. Identification of the isolates revealed that 31.2% were Pediococcus pentosaceus , 26.9% Lactococcus lactis , 18.6% Pediococcus acidilactici , 17% Lactobacillus brevis , and sporadic isolates of Leuconostoc mesenteroides , Lactobacillus plantarum , and Lactobacillus curvatus . Most of the strains did not produce gas from glucose and showed the capacity to produce lactic acid rapidly. Some 25% of the strains were able to degrade tributyrin (esterase activity), but none showed lipolytic activity against olive oil and pork fat. Only 3 strains of P. acidilactici showed weak proteolytic activity against myofibrillar or sarcoplasmic proteins. Also, the same strains showed antimicrobial activity against Listeria monocytogenes . Nine strains with the best properties were preselected and tested for biogenic amine production. The results showed that two of the strains, identified as P. acidilactici by polymerase chain reaction, had the potential to be further tested as starter cultures in pilot processing of Iberian sausages.     

Intraspecific diversity of Lactobacillus curvatus, Lactobacillus plantarum, Lactobacillus sakei, and Leuconostoc mesenteroides associated with vacuum-packed meat product spoilage analyzed by randomly amplified polymorphic DNA PCR

The intraspecific diversity of Leuconostoc mesenteroides, Lactobacillus curvatus, Lactobacillus sakei, and Lactobacillus plantarum was analyzed by randomly amplified polymorphic DNA (RAPD) PCR with universal primers M13 and T3. The study included 100 reference strains and 210 isolates recovered from two vacuum-packed Spanish meat products, fiambre de magro adobado and morcilla, previously identified by rDNA-restriction fragment length polymorphism profiles. The RAPD-M13 profiles identified isolates at species level in L. plantarum and L. mesenteroides, while RAPD-T3 provided profiles in L. sakei. The combination of RAPD-M13 and RAPD-T3 fingerprints revealed a total of 17 profiles in L. mesenteroides, 6 in L. sakei, 12 in L. plantarum, and 6 in L. curvatus. Of these, six profiles corresponding to L. mesenteroides and one corresponding to L. sakei were found in both products. The Shannon-Weaver diversity index (H'), calculated according to RAPD-M13 and RAPD-T3 profiles during storage, revealed that most profiles appeared only in single samplings in both products, indicating a high strain substitution rate during chilled storage of vacuum-packed meat products. When bloating appeared, only one profile corresponding to L. mesenteroides subsp. dextranicum was present throughout the storage period.     

Identification of lactic acid bacteria isolated from kimchi and studies on their suitability for application as starter culture in the production of fermented sausages

The aim of the investigation was to identify strains of lactobacilli coming from kimchi with properties suitable for use as starter cultures in sausage fermentation. A total of 31 strains of lactobacilli were isolated from kimchi on the 4-6th day of fermentation at 20°C using MRS agar plates and identified on the basis of morphological, biochemical, and physiological characteristics. The isolates were identified as Leuconostoc mes.mes./dent (12.9%), Lactobacillus curvatus (9.7%), Lactobacillus brevis (35.5%), Lactobacillus sake (25.8%), and Lactobacillus plantarum (16.1%). Thus, 51.6% of the isolates were homo-fermentative or facultative hetero-fermentative bacteria and the rest (48.4%) were hetero-fermentative bacteria. Among them L. brevis, L. curvatus, L. plantarum, and L. sake were investigated for their growth profile and metabolism characteristics in the fluid (submerged) model-medium modified according to the special conditions of fermented sausages. Relatively good growth properties were found for L. brevis, L. plantarum, and L. sake with maximum numbers of 8.18, 8.51 and 8.17cfu/ml, respectively, whereas L. curvatus could not adapt to the special environmental conditions. Regarding souring properties, L. brevis showed little ability to decrease pH, whereas L. curvatus, L. plantarum, and L. sake showed relatively good acidifying properties. According to the results of glucose fermentation and its products, only L. plantarum exhibited homo-fermentative characteristics. As a result only L. plantarum among the isolates from kimchi had an ability to adapt to the complex environment of fermented sausage, which will thereby allow them to act as starter cultures and natural preservatives in sausage production.     

PCR-DGGE分析东北自然发酵酸菜中乳酸菌多样性

为了探明传统的自然发酵白菜中乳酸菌的多样性及其优势乳酸菌群,试验主要采用变性梯度凝胶电泳法(denaturing gradient gel electrophoresis,DGGE),对5份采自我国东北地区利用传统方法制作的自然发酵酸菜样品中的乳酸菌多样性进行分析。结果表明,5份传统发酵酸菜样品共鉴定出了9个乳酸菌种,呈现出丰富的多样性。其中,植物乳杆菌、短乳杆菌、清酒乳杆菌和弯曲乳杆菌是酸菜样品的优势菌群,此外,还发现了片球菌、乳酸乳球菌、Hammesii乳杆菌和Odoratitofui乳杆菌,而Hammesii乳杆菌和Odoratitofui乳杆菌在此前文献报道中利用传统方法没有分离到。     

腌菜中细菌多样性研究及乳酸菌分离鉴定

以恩施地区采集的腌菜为研究对象,采用高通量测序技术（HTS）与变性梯度凝胶电泳（DGGE）技术相结合的方法对其中所 含细菌和乳酸菌多样性进行研究,并采用平板稀释涂布法对乳酸菌进行分离鉴定。结果表明,腌菜样品中含量最高的优势细菌门 为硬壁菌门（Firmicutes）,其平均相对含量高达97.09%。优势细菌属为Lactobacillus、Weissella、Leuconostoc、Vibrio、Pseudomonas、 Psychrobacter和Flavobacterium,其中Lactobacillus的平均相对含量高达82.37%;通过变性梯度凝胶电泳技术从样品中检测出的乳酸 菌 有 Lactobacillus sakei、Lactobacillus insicii、Lactobacillus plantarum subsp. argentoratensis、Lactobacillus plantarum subsp. plantarum 和 Lactobacillus acetotolerans;分离鉴定及保藏的乳酸菌中有L. plantarum 7株,L. alimentarius 2株,L. curvatus和L. brevis各1株以及L. sakei 4株。 由此可见,虽然Lactobacillus为腌菜样品中的优势菌,但乳酸杆菌的构成在样品间存在一定的差异。     

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.     

Isolation of bacteriocinogenic Lactobacillus plantarum strains from ben saalga, a traditional fermented gruel from Burkina Faso

A collection of lactic acid bacteria isolated from ben saalga, a traditional fermented gruel from Burkina Faso, was screened for bacteriocin production. Seven isolates were selected for their broad antimicrobial spectra, which overall included strains of Bacillus cereus, Bacillus licheniformis, Enterococcus faecalis, Listeria innocua, Listeria monocytogenes, Staphylococcus aureus, Escherichia coli and Salmonella enterica. Cluster analysis of RAPD-PCR patterns revealed that six of the isolates represent different strains. The six selected strains were identified as Lactobacillus plantarum by 16S rDNA sequencing, species-specific PCR and multiplex PCR of the recA gene. PCR amplification revealed the presence of genes of the plantaricin cluster described in L. plantarum C11. Among them, strain 5.2.2 carried the largest number of genes from this cluster.     

抗幽门螺旋杆菌乳酸菌的筛选与益生特性评价

目的:从发酵食品、乳制品和婴儿粪便中筛选具有拮抗幽门螺旋杆菌活性且益生特性良好的乳酸菌。方法:从175份样品中筛选获得39株具有良好遗传稳定性的疑似乳酸菌株,采用体外抑菌圈法和尿素酶活性试验筛选出能抑制幽门螺旋杆菌生长的9株菌,并对其中活性最强的菌株GQ1702和D1进行16S rDNA鉴定。然后,进一步评价其耐酸、耐胆盐、抗生素耐药性、抑菌性能、表面疏水性及自凝集性等益生特性。结果:筛选得到两株拮抗幽门螺旋杆菌的乳酸菌:瑞士乳杆菌GQ1702（Lactobacillus helveticus GQ1702）和植物乳杆菌D1（Lactobacillus plantarum D1）,其发酵产物的抗幽门螺旋杆菌活性分别相当于浓度为0.038 mg/mL和0.034 mg/mL甲硝唑的抑制率。在pH为3的环境中培养3 h后的存活率均大于70%,在含0.3%胆盐环境中培养3 h后存活率分别为0.47%和1.08%,疏水作用力分别为25.10%和11.79%,自凝集率在24 h后分别达到75.53%和68.61%,与幽门螺旋杆菌的交互凝集率在24 h分别达到61.18%和57.25%。对常见抗生素（氯霉素、青霉素、红霉素、氨苄青霉素和头孢唑林）不耐药,对4种肠道致病菌（金黄色葡萄球菌、单核细胞增生李斯特菌、大肠杆菌O157:H7、鼠伤寒沙门氏菌）均具有抑制作用。结论:本试验筛选到具有拮抗幽门螺旋杆菌能力的植物乳杆菌D1和瑞士乳杆菌GQ1702,可为功能性乳制品的开发及研究提供菌株。     

Diversity and probiotic potentials of lactic acid bacteria isolated from gilaburu,a traditional Turkish fermented European cranberrybush (Viburnum opulus L.) fruit drink

The aim of the present study was to characterize lactic acid bacteria (LAB) strains isolated from traditional fermented gilaburu fruit juice and their probiotic potential. The LAB counts of the fermented gilaburu fruit juice were in the range of 3.92–8.30 log cfu/g. Total of 332 isolates belonging to Lactobacillus and Leuconostoc species were characterized from traditional fermented gilaburu juice by genotypic methods. It was also determined that the major LAB strains belong to Lactobacillus plantarum (173 isolates), Lactobacillus casei (52 isolates) and Lactobacillus brevis (24 isolates), while Lactobacillus buchneri, Lactobacillus parabuchneri, Lactobacillus pantheris, Leuconostoc pseudomesenteroides and Lactobacillus harbinensis were the least in isolated LAB strains. In terms of the probiotic potentials, Lb. plantarum strains were able to grow at pH 2.5, but 3 of Lb. casei strains, one of each Lb. brevis and Lb. buchneri strains could not grow at the same pH. All selected LAB stains were resistant to bile salt at ≤ 0.3% concentration. While all the LAB species grew at 15 °C, two Lactobacillus hordei strains could also grow at 45 °C. The highest cell hydrophobicity degrees were for Lb. casei (G20a) and Lb. plantarum (G19e) as 87.5 and 86.0%, respectively. Listeria monocytogenes and Bacillus cereus were the most sensitive bacteria against the selected LAB strains, while Escherichia coli and Staphylococcus aureus were the most resistant. Again all the isolated LAB species were resistant to three antibiotics; kanamycin, streptomycin and vancomycin. Characterization and probiotic potentials of the LAB isolated from fermented gilaburu (Viburnum opulus) juice were studied first time, and further research needs to be done on their behaviors in similar food formulations as a probiotic.     

Production of bacteriocin-like metabolites by lactic acid cultures isolated from sucuk samples

A total of 51 sucuk samples, obtained from different regions of Turkey, were examined for lactic acid bacteria (LAB) strains producing putative bacteriocin-like metabolites. For detection of antagonistic activity, agar spot and well diffusion assay tests were used. Lactobacillus sake Lb790, Listeria monocytogenes Li6, Staphylococcus aureus St44 and Escherichia coli NRRL B-3704 were used as indicator organisms. Strains having antimicrobial activity were also tested against Clostridium perfringens 4TTK, Clostridium botulinum type A. 7948, Bacillus cereus NRRL B-3711, Micrococcus luteus NRRL B-4376 and Yersinia enterocolitica type 103. 57 of 424 isolates from sucuk samples were putative producers of bacteriocin-like metabolistes and were identified as the following: 19 Lactobacillus plantarum, 4 L. curvatus, 4 Pediococcus pentosaceus, 3 P. acidilactici, 2 L. pentosus, 2 L. sake, 1 L. delbrueckii, 1 L. rhamnosus and 21 unidentified Lactobacillus spp. P. pentosaceus 416, P. acidilactici 413, 419 and 446, L. curvatus 348, L. plantarum 452 and 495, and Lactobacillus spp. 411 strains have the best potential for use as sucuk starter cultures.     

大庆自然发酵酸菜中乳酸菌的分离鉴定及耐酸菌株初步筛选

从黑龙江大庆地区采集7份采用传统方法制作的自然发酵酸菜发酵液,从中分离和筛选出14株乳酸菌疑似菌株,提取其16S rDNA,并经测序、同源性分析和系统发育树构建等方法,对其属种进行鉴定。初步筛选出在pH值为2.5、3.0和3.5的酸性条件下均能够生长的耐酸菌株6株,并进一步利用活菌计数法得出菌株在pH3.0条件下的存活率。结果表明：14株菌株均为乳酸菌,其中4株为弯曲乳杆菌（HD12-1、HD13-5、HD14-1和HD15-1）,1株为短乳杆菌（HD18-2）,3株为清酒乳杆菌（HD12-2、HD13-1和HD16-5）,1株为肠膜明串珠菌（HD18-3）,5株为植物乳杆菌（HD14-3、HD15-2、HD16-2、HD17-3和HD17-4）,且筛选出pH 3.0条件下存活率在2%以上的6株菌株,分别为HD12-1、HD13-1、HD14-1、HD15-1、HD16-2和HD16-5。     

Evaluation of meat born lactic acid bacteria as protective cultures for the biopreservation of cooked meat products

In this study, 91 strains, originating from meat products, were subjected to a step-by-step screening and characterisation to search for potential protective cultures to be used in the cooked cured meat industry. Strains were first tested on their homofermentative and psychrotrophic character and salt tolerance. Secondly, the antibacterial capacities towards Listeria monocytogenes, Leuconostoc mesenteroides, Leuconostoc carnosum and Brochotrix thermosphacta were determined in an agar spot test. In total, 38% of the tested strains were inhibitory towards all indicator strains. However, 91%, 88% and 74% of the strains could inhibit, respectively, L. monocytogenes, B. thermosphacta and Leuc. mesenteroides. Finally, 12 strains, with the highest antibacterial capacities, were evaluated on their competitive nature by comparing their growth rate, acidifying character and lactic acid production at 7 degrees C under anaerobic conditions in a liquid broth. All 12 strains, except for a bacteriocin producing Lactobacillus plantarum strain and the lactocin S producing Lactobacillus sakei 148, combined a fast growth rate with a deep and rapid acidification caused by the production of high levels of lactic acid. The 12 selected strains were then further investigated for their growth capacity on a model cooked ham product to establish whether the presence of these cultures on the ham did not negatively influence the sensory properties of the ham. All strains grew in 6 days at 7 degrees C from a level of 10(5)-10(6) to 10(7)-10(8) cfu/g and again the bacteriocin producing L. plantarum strain was the slowest growing strain. As the glucose level of the model cooked ham product was low (0.09+/-0.03%), growth of the putative protective cultures resulted in glucose depletion and a limited lactic acid production and accompanying pH decrease. Cooked ham inoculated with isolates 13E, 10A, 14A (all three identified as L. sakei subsp. carnosus by SDS-PAGE) and with strains L. sakei 148 (LS5) and L. sakei subsp. carnosus SAGA 777 (LS8) were not rejected by the sensory panel at the 34th day of the vacuum packaged storage at 7 degrees C. Therefore, these strains could have potential for the use as protective culture in cooked meat products.     

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.     

Lactobacillus role during conditioning of refrigerated and vacuum-packaged Argentinean meat

The role of Lactobacillus strains with bioprotective and technological potential on raw beef during 15days of storage under vacuum at 7°C was investigated. The assayed strains were able to grow on the meat, Lactobacillus curvatus CRL705 and Lactobacillus sakei 23K showing the highest competitiveness. A net increase of amino acids was determined in inoculated samples when compared to the control, this being maximal for Lactobacillus plantarum CRL681. Although an important endogenous activity of meat sarcoplasmic proteins was observed, the disappearance of protein bands and the generation of a new one were detected as a consequence of Lactobacillus growth. A synergistic effect of Lactobacillus in combination with the muscle proteolytic enzyme complex can be suggested. From the studied strains, the bacteriocin producer L. curvatus CRL705 may be considered as a good candidate to contribute to meat ageing by means of small peptides and free amino acids generation while improving shelf life.     

生物保鲜乳酸菌的筛选及抑菌物质性质的初步研究

从收集保藏的乳酸菌中,筛选出对大肠杆菌、荧光假单胞菌、蜡状芽孢杆菌单核细胞增生李氏杆菌和金黄色葡萄球菌这些常见食品腐败菌都表现出良好抑菌效果,具有生物保鲜价值的5株乳酸菌,其中3株植物乳杆菌(ACCC10171、ACCC11095和ACCC11118),1株干酪乳杆菌(ACCC11052)和1株类干酪乳杆菌(ACCC11050)。在排除酸性末端产物和过氧化氢的干扰后,用蛋白酶K等多种蛋白酶处理这5株菌的代谢产物粗品,其抑菌活性都受到一定影响,甚至完全丧失抑菌活性,因此可以推断其代谢物质中含有对蛋白酶敏感的细菌素类物质。     

Molecular typing techniques to characterize the development of a lactic acid bacteria community on vacuum-packaged beef

The development of a community of lactic acid bacteria from vacuum-packaged beef was investigated during a 6-week storage trial at 2 degrees C. The lactic acid bacteria population was monitored by using molecular techniques to identify a random sample of isolates at biweekly intervals during the storage trial. The polymerase chain reaction and a randomly amplified polymorphic DNA technique were used to identify and distinguish populations of lactic acid bacteria that developed during the storage trial. At week 0, the population of lactic acid bacteria was 3.5 log cfu/120 cm2 and by week 6, the population reached a maximum of 7.6 log cfu/120 cm2. A sampling from the week 0 population indicated a mixed community of Lactobacillus curvatus, Lactobacillus sakei and Leuconostoc spp. However, the sampling from week 6 indicated the population composition had changed to one where a single Leuconostoc strain predominated. This strain demonstrated antagonism towards the growth of other lactic acid bacteria isolated during the study. Additionally, the strain inhibited the growth of foodborne pathogens Escherichia coli O157:H7 and Listeria monocytogenes. DNA sequence data from the 16S rRNA gene suggested that the isolate may be a Leuconostoc gelidum strain.     

Characterisation of naturally fermented sausages produced in the North East of Italy

In the Friuli Venezia Giulia region, in the North East of Italy, a traditional fermented sausage is produced without the use of microbial starters. It is characterized at the end of the ripening period by accentuated acidity, slight sourness and elastic, semi-hard consistency. In this study, three fermentations, carried out in different seasons (winter, spring and summer) were followed analyzing the microbiological, physicochemical and sensory aspects of this product. The sausages were characterized by an important microbial activity of lactic acid bacteria and micro/staphylococci that resulted in a product with a final pH of about 5.6-5.7. An interesting aspect was the high number of fecal enterococci that can play an important role in the definition of the organoleptic profile of the final product. No Listeria monocytogenes, Salmonella spp. and Staphylococcus aureus were ever isolated from the raw materials or the fermented sausages during the maturation, underlining the safety of this product. The final water activity of the product was 0.91-0.92. One hundred and fifty lactic acid bacteria were isolated and identified by molecular methods to understand which species were more predominant in the product. Lactobacillus curvatus and Lactobacillus sakei were the most numerous (54 and 64 strains isolated, respectively) and they were the only species common to all three fermentations. A cluster analysis of the profiles obtained from these strains after RAPD-PCR highlighted a population distribution that was fermentation-specific.     

Screening of glutamate decarboxylase activity and bile salt resistance of human asymptomatic carriage, clinical, food, and environmental isolates of Listeria monocytogenes

Following consumption, stomach acidity is the first major barrier encountered by the food-borne pathogen Listeria monocytogenes. Analysis of low pH sensitivity and glutamate decarboxylase (GAD) acid resistance system of 14 isolates of L. monocytogenes carried asymptomatically by humans showed that levels of GAD activity were subjected to strain variation. Similar variations were observed for strains responsible for 18 cases of listeriosis, whereas in comparison, 13 strains isolated from food and food-processing plant environments showed lower GAD activity. Following survival of the stomach barrier, L. monocytogenes also has to resist bile salts encountered in the small intestines. Analysis revealed that all strains tested were able to grow in the presence of bile salts with concentrations as high as those encountered in the small intestines and had previously identified bile salt hydrolase (BSH) activity. Strain variation was observed but there was no relationship between the origin of the strains and the ability to degrade bile salts.     

Enhancing the antilisterial effect of Lactobacillus curvatus CWBI-B28 in pork meat and cocultures by limiting bacteriocin degradation

This work focused on Listeria monocytogenes growth inhibition and growth rebound in raw and cooked pork meat inoculated with Lactobacillus curvatus strains. During storage of raw meat homogenates in the presence of the bacteriocin-producing strain Lactobacillus curvatus CWBI-B28wt, the Listeria monocytogenes cfu count was initially reduced to an undetectable level, but a growth rebound occurred after two weeks, coinciding with loss of 70% of the bacteriocin activity present at the end of week 2. The Listeria growth rebound was suppressed when proteolysis of bacteriocin was countered by the absence of proteases (bacteriocin addition to cooked meat) or the presence of 1% soy flour (added to provide competing substrates). Further experiments confirmed that bacteriocin is sensitive to the action of proteolytic enzymes isolated from both Lactobacillus curvatus CWBI-B28wt and the meat matrix. Bacteriocin proteolysis thus emerges as a cause of Listeria growth rebound.     

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.