Aroma components from dried sausages fermented with Staphylococcus xylosus

Sausages with and without Staphylococcus xylosus were manufactured with four replicates. Antibiotics and a fungicide to inhibit growth of naturally occuring microorganisms were added to the control sausages. The volatile compounds from the sausages were collected and identified by gas chromatography/mass spectrometry, the effluent evaluated by sniffing and the sausages assessed by a trained panel. The study showed that sausages fermented with Staphylococcus xylosus contained several fragrant esters that were not found in control sausages without microbial growth. Control sausages had an unpleasant, rancid odour compared to sausages with Staphylococcus xylosus, indicating that the esterase activity of Staphylococcus xylosus or other microorganisms is very important in order to obtain the proper fermented sausage aroma. Although sausages with Staphylococcus xylosus contained the highest amounts of free fatty acids, it seemed to be of no importance to the aroma development. It is therefore questionable whether lipolytic activity of starter cultures has any influence on sausage flavour.     

Growth and aroma production by Staphylococcus xylosus, S. carnosus and S. equorum--a comparative study in model systems

A laboratory medium inoculated with 20 different Staphylococcus strains was prepared in accordance with a full factorial experimental design investigating the effect of temperature, pH, NaCl and glucose on growth. The 12 strains most suited to growth in a fermented meat environment were inoculated in sausage minces together with Pediococcus pentosaceus, incubated at 25 degrees C for 1 week and the produced aroma compounds collected. The data were analysed by multiple linear regression and partial least squares regression analysis. The results showed that increasing pH and temperature from 4.6 to 6.0 and 10 to 26 degrees C, respectively, increased growth of all strains with strong synergy between temperature and pH. Increasing salt concentration from 5% to 15% w/v decreased growth of most strains, but the effect of pH and temperature was much stronger than the effect of salt. Strains of S. carnosus were more salt tolerant than strains of S. equorum and S. xylosus, especially at high pH and temperature. Addition of glucose up to 0.5% w/v had no significant influence on growth of any of the strains. With regard to aroma production, species characteristics were detected. S. carnosus and S. xylosus were quite different regarding the overall aroma profiles, whereas the profiles of S. equorum lied somewhere in-between. Contrary to S. carnosus, S. xylosus and S. equorum did not produce 2-methyl-1-butanol. On the other hand, in particular, S. xylosus produced more 3-methyl-1-butanol. Except for one of the strains of S. equorum, S. xylosus and S. equorum formed more diacetyl, 2-butanone and acetoin and also more of the methyl-branched ketones arising from degradation of leucine, isoleucine and valine. S. carnosus produced more methyl-branched aldehydes, acids and corresponding esters from leucine, isoleucine and valine compounds that have been correlated with fermented sausage maturity in former studies. S. equorum produced the least of the methyl-branched aldehydes.     

Effects of muscle shortening and proteolysis on Warner-Bratzler shear force in beef longissimus and semitendinosus

The effect of the addition of the fungal protease EPg222 on the sensory characteristics of dry fermented sausage "salchichón" ripened with commercial starter cultures was investigated. Sausages were prepared with purified EPg222 and Staphylococcus carnosus, Staphylococcus xylosus, and Lactobacillus sakei as starter cultures, ripened for 145 days and compared with a control batch only inoculated with the starter cultures. Dry fermented sausages ripened with EPg222 and starter cultures showed higher amount of NPN and volatile compounds derived from amino acid catabolism, than control ripened only with starter cultures. Several branched aldehydes, acids and alcohols such as 2- and 3-methylbutanoic acid and 2-methylpropanol were detected only in enzyme treated samples. Sensory analysis reflected higher values for aroma intensity of sausages treated with EPg222 and lower values of hardness than control. The effect of EPg222 may be of great interest to improve sensory characteristics of dry fermented sausages ripened with starter cultures.     

Proteolytic and lipolytic starter cultures and their effect on traditional fermented sausages ripening and sensory traits

In this study, three starter formulations including Lactobacillus curvatus and Staphylococcus xylosus strains selected in vitro on the basis of their lipolytic and proteolytic activities were employed for the manufacture of traditional fermented sausages of southern Italy. Microbial population, proteolysis, lipolysis, changes in free amino acids (FAA) and free fatty acids (FFA) and development of characteristic taste and flavor of the final product were investigated. Proteolysis and lipolysis were observed in sausages inoculated with proteolytic and lipolytic S. xylosus coupled with L. curvatus, while the sausage started with only S. xylosus without lactobacilli was identical to the non-inoculated control, indicating that the proteolysis could be due to both microbial activity and endogenous proteases activated by the decrease in pH. The statistical analysis applied to the instrumental and sensory data showed that there was an effect of the starter used on the characteristics of the sausage obtained. In particular, the control samples showed very close features different from the sausages obtained by adding starter cultures. Finally, analyzing the sensory parameters the sausages ripened without starter addition and those started without the L. curvatus AVL3 showed similar features indicating an influence of the presence of the lactobacilli on the final organoleptic quality of the sausages. An appropriate choice of a combination of strains in a starter formulation is fundamental to obtain products of the expected quality.     

The influence of Debaryomyces hansenii and Candida utilis on the aroma formation in garlic spiced fermented sausages and model minces

The influence of the yeast starter cultures Debaryomyces hansenii and Candida utilis on fermented meat aroma was studied in model minces and in commercial-type fermented sausages. Volatile compounds from model minces and sausages were collected using diffusive and dynamic headspace sampling respectively and were identified by gas chromatography/mass spectrometry (GC/MS). A triangle test was carried out on the sausages to detect whether the yeast influenced the sausage odour. C. utilis demonstrated high metabolic activity in the model minces, producing several volatile compounds, in particularly esters. C. utilis also seemed to ferment the amino acids valine, isoleucine and leucine into compounds important for the aroma of sausages. D. hansenii on the contrary, had very little effect on the production of volatile compounds in the model minces. In the sausage experiment both yeast cultures died out before the ripening process ended and the sensory analysis showed only a slight difference between the sausages. A fungistatic test of the garlic powder added to the sausages indicated that garlic inhibits the growth of the yeast starter cultures.     

微生物酶与肉组织酶对干发酵香肠中游离氨基酸的影响

以戊糖乳杆菌31-1和木糖葡萄球菌为发酵剂生产干发酵香肠,就发酵剂对干发酵香肠成熟过程中游离氨基酸的影响进行了研究,同时对干香肠在成熟过程中微生物和pH值的变化进行了分析。研究结果表明,微生物的代谢活动能增加干发酵香肠中游离氨基酸的绝对含量,但相对组织酶来说,微生物酶对游离氨基酸的解离能力要小得多,说明在干发酵香肠的成熟过程中,组织酶对蛋白质的水解起主要作用。但研究结果同时也表明,微生物对个别游离氨基酸的变化具有最大的影响。     

发酵剂对发酵香肠食用品质的影响及其在不同直径香肠中的应用

以商业发酵剂（木糖葡萄球菌＋戊糖片球菌）为对照组,不同发酵剂组合（木糖葡萄球菌＋副干酪乳杆菌、木糖葡萄球菌＋戊糖片球菌＋副干酪乳杆菌）为实验组,通过对发酵香肠水分含量、pH值、水分活度、色泽、质地、风味和感官品质等指标进行测定,确定最佳发酵剂及适宜发酵的香肠直径。结果表明：相较于商业发酵剂,木糖葡萄球菌与副干酪乳杆菌组合发酵的香肠总体可接受性相对较高,且庚醛、1-辛烯-3-醇、乳酸乙酯、戊酸乙酯、癸酸乙酯、2-甲基丙酸乙酯等愉悦风味物质为该组独有,其特征主要表现为更加浓郁的清新味、甜香味、果香味和花香味;适宜的直径（21 mm）、水分含量（（25.40±0.00）%）和硬度（（2 812.46±767.93）g）以及相对较高的pH值（pH 5.57±0.02）是该组发酵香肠口感显著高于其他2 组的重要原因。因此,木糖葡萄球菌与副干酪乳杆菌组合发酵的小直径香肠食用品质最佳。     

复合发酵剂和天然香辛料对羊肉发酵香肠品质特性的影响

通过接种复合发酵剂和添加天然香辛料,采用相同的工艺条件生产4组羊肉发酵香肠,1组为对照组,2组为接种发酵剂组,3组为发酵剂+黑胡椒组,4组为发酵剂+黑胡椒+孜然组。分别测定4个组羊肉发酵香肠在发酵成熟和贮藏过程中pH值、Aw值、水分含量及微生物的变化。结果表明:在发酵结束(即第1.5天)时,试验组和对照组的pH值、Aw值和水分含量均呈下降趋势,试验组的pH值迅速降到最低值(即5.0以下),与对照组差异显著(P<0.05);在干燥成熟过程中,试验组的pH值、Aw值和水分含量迅速下降,添加香辛料组的pH值迅速回升,同时乳酸菌数维持在108cfu/g以上水平,高于对照组,细菌总数却低于对照组;在贮藏过程中,试验组的pH值和水分含量基本保持不变,且均低于对照组,而Aw值始终呈下降趋势,细菌总数小于对照组。上述结果表明,接种复合发酵剂和添加天然香辛料能迅速降低pH值、Aw值和水分含量,有效抑制腐败菌和致病菌的生长,提高羊肉发酵香肠的品质和保藏性。     

Occurrence and significance of streptococci in fermented Italian type dry sausage

Commercial cultures used in Canada for the manufacture of Italian dry sausage were examined to determine their microbial composition and suitability for low temperature (less than or equal to 20 degrees C) meat fermentations. Temperature optima in both laboratory media and commercial meat mixtures were generally too high to allow these cultures to be of substantial advantage in this application. In addition, media used currently for the enumeration of streptococci and related organisms from fermented meat products were found to be inadequately specific and often required confirmatory inspection of colonies by conventional phase contrast microscopy. Streptococci were isolated from Italian dry sausage manufactured commercially with and without added starter cultures. Streptococci persisted in sausages produced by both techniques with slightly higher numbers present in starter-acidulated sausages. About 55.5% of the 312 streptococci studied were enterococci (Lancefield's Group D). Streptococci were found in several samples of commercial starter cultures but it was felt that elevated ripening temperatures used for sausage manufactured by the starter-mediated process and meat handling practices were more important factors influencing streptococci recovery from sausage material.     

Volatile compound generation in dry fermented sausages by the surface inoculation of selected mould species

The effect of the inoculation of dry fermented sausage surface with an atoxigenic, proteolytic and lipolytic strain of Mucor racemosus, Penicillium aurantiogriseum and Penicillium camemberti on the volatile composition was studied. The analysis of the headspace volatile compounds using gas chromatography/mass spectrometry enabled the identification of 55 volatiles. The study showed that every mould species produced a different volatile profile which was also different from that of the control sausages. Compounds derived from amino acid catabolism, i.e. branched aldehydes and the corresponding alcohols, were produced in higher amounts in sausages inoculated with Penicillium spp. On the other hand, volatiles coming from the microbial esterification were related to sausages inoculated with M. racemosus. The development of the fungal mycelia on the sausage surface protected lipids from oxidation, thus giving rise to fewer lipid oxidation products in the inoculated sausages.     

中式发酵香肠用发酵剂混合菌种的研究

将玫瑰色微球菌、干酪乳杆菌、植物乳杆菌、戊糖片球菌、嗜酸乳杆菌、木糖葡萄球菌采用正交试验法组合为 9组发酵剂 ,考察了发酵肠的 pH值、水分含量、乳酸菌活菌数、感官品质等的变化规律。结果表明 ,混合球菌更适合于作为发酵肠的发酵剂。经优化组合 ,筛选出了由葡萄球菌、干酪乳杆菌、玫瑰色微球菌、嗜酸乳杆菌、戊糖片球菌等混合菌制备的发酵剂 ,是符合发酵肠GMP要求的快速发酵剂 ,且经济合理 ,并赋予产品良好的风味。发酵条件 :37℃ ,RH >80 % ,2 0h。     

Isolation and technological properties of coagulase negative staphylococci from fermented sausages of Southern Italy

The aims of this study were to characterize the population of Micrococcaceae in different types of fermented sausages of Southern Italy and to determine the technological properties of Staphylococcus strains in order to evaluate the suitability of selected strains as starter cultures in the processing of dry fermented pork sausages. Ninety-six strains were studied to evaluate nitrate reductase, proteolytic, lipolytic and antioxidant activities as well as growth ability at different temperatures, pH's and NaCl concentrations. All the strains were classified as Staphylococcus except for one isolate assigned to Kocuria spp. The species most often isolated were S. saprophyticus, S. xylosus and S. equorum, although they were not equally distributed within the different sausages. Other species isolated were, in descending order of abundance, S. succinus, S. warneri, S. lentus, S. vitulus, S. pasteuri, S. epidermidis, and S. haemolyticus. In general, the S. xylosus strains exhibited the best technological properties that would make them eligible as good starter cultures for fermented meat products. However, strains belonging to other species also showed good technological properties. Finally, all strains grew at 10, 15 and 20 °C, in the presence of 10% and 15% of NaCl and at pH 5.0 and 5.5. The results showed that it is possible to formulate a broad variety of staphylococcal starter cultures, adaptable to different technological conditions and sausage manufacture practices.     

Identification of potential probiotic starter cultures for Scandinavian-type fermented sausages

Potential probiotic cultures suitable as starter cultures for the Scandinavian-type fermented sausages were identified among strains well-adapted to fermented meats as well as strains originating from a culture collection. From 15 different fermented meat products, 22 strains were isolated as dominant non-starter lactic acid bacteria (NSLAB). The isolates were identified by RAPD, API and sequence analysis of 16S rRNA and showed to be five strains of Lactobacillus sakei, five strains of Lactobacillus farciminis, five strains belonging to the group of Lactobacillus plantarum/pentosus, four strains of Lactobacillus alimentarius, two strains of Lactobacillus brevis and one strain of Lactobacillus versmoldensis. Heterofermentative strains as well as strains not growing at 37 degrees C and not lowering pH below 5.1 in a meat model were excluded leaving 9 strains for further studies. These strains together with 19 strains from a culture collection were evaluated by in vitro methods including survival upon exposure to pH 2.5 or 0.3% oxgall and adhesion to the human colon adenocarcinoma cell line Caco-2 as well as antimicrobial activity against potential pathogens. Strains that fulfilled all the probiotic criteria and showed to be fast acid producers in a meat model included three strains belonging to the group of Lb. plantarum/pentosus (MF1291, MF1298, MF1300) which originated from the dominant NSLAB of fermented meat products. MF1291 and MF 1298 were further identified as Lb. plantarum and MF1300 as Lb. pentosus. The three strains were all successfully applied as starter cultures for the production of fermented sausage. The viable count at the end of the processing period reached high cell numbers (4.7x10(7)-2.9x10(8) cfu/g) and pH of the sausages decreased to pH 4.8-4.9 without any flavour deviation compared to sausage fermented by a commercial meat starter culture.     

人工发酵剂对湘西腊肠微生物菌群和理化特性的影响

本文从动态角度探讨自然发酵、戊糖片球菌发酵、木糖葡萄球菌发酵和混合菌发酵腊肠在发酵过程中五种菌相(细菌总数、乳酸菌、葡萄球菌、酵母菌和肠细菌)和理化特性的阶段性变化。微生物数据显示,四组腊肠初始肠细菌数量约5.30 logCFU/g,而发酵结束,添加了戊糖片球菌腊肠组的肠细菌数量约1.00 logCFU/g,没有添加该菌腊肠组的肠细菌数量约3.30 logCFU/g,说明戊糖片球菌能有效抑制肠细菌的生长,保证了产品的安全性和稳定性;发酵初期,腊肠p H值迅速降5.30以下,到发酵中期,腊肠p H开始回升;在发酵过程中,戊糖片球菌腊肠组的TBARS(硫代巴比妥酸)值显著(p0.05)高于其他三组腊肠,而其它理化特性无显著差异。从数据分析可知,腊肠的发酵剂能有效改善其卫生质量,而对腊肠的p H值、AW和亚硝酸盐残留量没有显著影响。     

不同乳酸菌发酵剂对发酵红肠品质的影响

为研究乳酸菌发酵对红肠品质的影响,将发酵技术应用于本无发酵工艺的红肠制品中,筛选出能够提高红肠品质的乳酸菌发酵剂。分别将常应用于发酵肉制品的7 种商业乳酸菌发酵剂（木糖葡萄球菌＋戊糖片球菌（THM-17）、木糖葡萄球菌＋清酒乳杆菌＋类植物乳杆菌（PRO-MIX5）、木糖葡萄球菌＋肉葡萄球菌＋清酒乳杆菌（WBL-45）、木糖葡萄球菌＋戊糖片球菌＋植物乳杆菌（VHI-41）、木糖葡萄球菌＋戊糖片球菌＋植物乳杆菌（SHI-59）、肉葡萄球菌＋木糖葡萄球菌（WBX-43）和戊糖片球菌＋木糖葡萄球菌＋肉葡萄球菌＋乳酸片球菌（VBM-60））及8 种单菌（弯曲乳杆菌、戊糖乳杆菌、清酒乳杆菌-1、戊糖片球菌、木糖葡萄球菌、肉葡萄球菌、清酒乳杆菌-2、植物乳杆菌）以107 CFU/g的接种量接种至腌制后的肉馅中,拌馅灌肠后于35 ℃、80%湿度条件下发酵12 h,取样测定发酵后样品的乳酸菌数和细菌总数,再经干燥、蒸煮、烟熏、烘烤制得成品,测定其感官、pH值、色差、质构、亚硝酸盐、硝酸盐、生物胺及N-亚硝胺含量等指标。结果表明：15 种发酵剂中以木糖葡萄球菌和植物乳杆菌2 种乳酸菌发酵剂应用效果较好,所制得产品pH值分别为5.26和5.04,色泽美观,弹性适中,亚硝酸盐残留量（10.84、10.13 mg/kg）低,可显著抑制N-亚硝胺的形成（N-二甲基亚硝胺含量分别为1.29、2.51 μg/kg）,生物胺总量较低。由此说明,木糖葡萄球菌和植物乳杆菌能够显著提高红肠产品的安全品质。     

不同原料肉对发酵香肠食用品质的影响

选取3种冷鲜肉(黑土猪肉、阳光猪肉、三元猪肉),采用商业发酵剂(木糖葡萄球菌+戊糖片球菌)接种发酵的方式制备符合中国消费者口味的切片即食型发酵香肠,分析不同原料肉对发酵香肠理化特征、风味及感官特性的影响.结果表明:3种原料肉制备的发酵香肠总体可接受度良好,光泽度高、色泽暗红(红度值8.62～9.85)、组织紧密、pH?...     

腌腊肉制品中乳酸菌的筛选鉴定及其在腊肠中的应用

为筛选适合传统腌腊肉制品的优良乳酸菌菌株,从多种农家自制传统腌腊肉制品中分离纯化出9株优势乳酸菌。通过发酵特性筛选,得到一株性状优良菌株10M-7,并制备该菌株的干粉发酵剂,以未接种发酵剂腊肠为对照,分析此发酵剂对腊肠感官品质和微生物变化的影响。结果表明,10M-7菌株具有良好的产酸特性和抑菌性能。根据形态学、生理生化特征和16S rRNA序列分析,鉴定其为植物乳杆菌,采用冷冻干燥法制备纯种发酵剂,并制作人工发酵腊肠。发酵剂组pH值在初期便迅速下降,且始终低于对照组;发酵剂组乳酸菌迅速生长繁殖,且葡萄球菌和大肠杆菌数量与对照组相比明显降低。感官评价表明,当添加量为10~4CFU/g原料肉时,能够很好地保持和改善产品风味,使产品整体感觉更好。     

Generation of flavour compounds in fermented sausages-the influence of curing ingredients, Staphylococcus starter culture and ripening time

The volatile profiles of fermented sausages made with either Staphylococcus xylosus or Staphylococcus carnosus starter cultures were studied with regard to the influence of salt concentration, ripening time and three different combinations of curing ingredients-nitrate, nitrite or nitrite/ascorbate. Emphasis was laid on volatile compounds originating from degradation of branched-chain amino acids. Volatile compounds were collected using dynamic headspace sampling and were identified by gas chromatography/mass spectrometry (GC/MS). Development in water activity, water loss and pH was monitored throughout maturation. Curing salts had a pronounced effect on the level of volatile compounds. In particular, curing with nitrate instead of nitrite resulted in a striking difference. Generally, nitrate increased the level of volatile compounds compared to nitrite, whereas ascorbate had only a small influence. The concentration level of NaCl had a considerable effect on the amount of volatile compounds but the effect was highly related to the ripening stage. Most compounds, but not all, increased in concentration as ripening proceeded. Major differences in the development of volatile compounds were observed depending on whether S. xylosus or S. carnosus were used as starter culture. In particular the effects of nitrate was much more predominant in the sausages made with S. carnosus than S.xylosus.     

Freezing of meat raw materials affects tyramine and diamine accumulation in spontaneously fermented sausages

Biogenic amine accumulation was studied in spontaneously fermented sausages (Fuet) manufactured from unfrozen-fresh meat (U-sausages) and frozen-thawed meat (F-sausages). The aim was to investigate whether the frozen storage of raw materials affects the microbial composition and its aminogenic activity during sausage fermentation. Tyramine was the major amine in all sausages. Although the final levels were similar, tyramine accumulated more rapidly in F-sausages, which contained putrescine as the second amine. By contrast, U-sausages accumulated much more cadaverine than putrescine. F-sausages showed a slightly lower pH and free amino acid content as well as higher counts of technological flora (lactic acid and gram positive catalase positive bacteria) and lower counts of enterobacteria. Therefore, to freeze the meat raw materials for few days before sausage manufacture could be a useful practice, especially for the artisan fermented sausages (without starter), because it helps to reduce enterobacteria development and cadaverine production.     

微生物酶与肉组织酶对干发酵香肠中游离脂肪酸的影响

以戊糖乳杆菌31-1和木糖葡萄球菌为发酵剂生产干发酵香肠,就发酵剂对干发酵香肠成熟过程中游离脂肪酸的影响进行了研究。研究结果表明,在灌肠后的1周内,脂肪的水解主要是由组织酶引起的,微生物对脂肪微弱的水解能力主要表现在干发酵香肠成熟的后期,并且发酵剂不能改变发酵香肠中游离脂肪酸的变化模式。