Development of a PCR-culture technique for rapid detection of yeast species in vacuum packed ham

A rapid assay for detection of yeast species in vacuum packed ham has been developed based on the polymerase chain reaction (PCR) coupled to a 24 h pre-enrichment at 25 °C. DNA was isolated from yeast inoculated ham samples and amplified using primers specific for the 18S rRNA gene sequences of yeasts. A detection limit of 10(2) CFU/cm(2) was achieved following enrichment of samples experimentally inoculated with three yeast species frequently associated with meat products spoilage: Debaryomyces hansenii, Yarrowia lipolytica, and Kluyveromyces marxianus. Likewise, commercial sliced and vacuum packed ham samples were analysed using the PCR-culture technique. The results obtained in this work show that PCR amplification of a conserved region of the 18S rRNA gene in the yeast species could be potentially used as a rapid tool for detection of low levels of viable spoilage yeasts in meat products.     

Application of a multiplex PCR for the simultaneous detection of Escherichia coli O157:H7, Salmonella and Shigella in raw and ready-to-eat meat products

Escherichia coli O157:H7, Salmonella and Shigella might contaminate similar types of meat products and cause deadly diseases in humans. Traditional microbiological analyses to detect these pathogens are labor-intensive and time-consuming. The objective of this study was to apply a multiplex PCR for simultaneous detection of the pathogenic bacteria in certain raw and ready-to-eat meat matrices. The tested samples had aerobic plate counts ranging from non-detectable, in chicken nuggets and salami, to 8.36log(10)CFU/g in ground pork. The pH of homogenates spanned from 6.86, in ground beef, to 7.17 in salami. Following a 24-h enrichment, the multiplex PCR assay could concurrently detect the three pathogens at 0.2log(10)CFU/g in ground beef, roast beef, beef frankfurters, chicken nuggets, salami and turkey ham, and 1.2log(10)CFU/g in ground pork. This multiplex PCR offers an efficient microbiological tool for presumptive detection of E. coli O157:H7, Salmonella and Shigella in meat.     

Dynamics and characterization of yeasts during ripening of typical Italian dry-cured ham

The evolution of the yeast population during manufacturing and ripening of dry-cured Parma ham was investigated. Contamination levels ranged from 10(5) to 10(7) cfu/g on muscle surface, 10(4) to 10(6) cfu/g on covering fat and exceeded 10(7) cfu/g on spreadable fat mince ("sugna"). Two hundred and sixty one yeast isolates underwent identification test, showing that the predominant species of yeast population during the whole maturing process were Debaryomyces hansenii, Candida zeylanoides, Debaryomyces maramus, and to a lesser extent, Candida famata and Hyphopichia burtonii. The species Candida catenulata, Candida guilliermondii, Candida edax and other genera like Cryptococcus and Wingea were occasionally found. The yeast counts and species distribution changed according to the stage of processing and to the ham sampling location. At the end of the cold phase, the washing procedure was effective in lowering the yeast count in muscle and fat surface layers, but during the next ageing stages, yeast colonization of unskinned ham muscle increased again, though species distribution changed if compared to previous manufacturing phases. The ripening steps taken into account from the end of the cold phase to the final outcome, were always characterized by more than one yeast species, suggesting that yeasts other than Debaryomyces spp. could play a remarkable role on the sensory and safety properties of typical Italian dry-cured ham.     

Literature compilation of volatile N-nitrosamines in processed meat and poultry products - an update

ABSTRACT

A compilation of volatile N-nitrosamine levels in processed (e.g., cured, canned, smoked) meat and poultry products is presented. Over 1800 samples of processed meat products including bacon, ham, salami, sausage, and various other processed meat and poultry products have been examined for the presence of eight volatile N-nitrosamines. The database compiled from the literature is based on 25 references published for the period of 1985 to 2018 from 14 countries. N-nitrosodimethylamine (NDMA), N-nitrosopiperidine (NPIP), and N-nitrosopyrrolidine (NPYR), are the most frequently identified volatile N-nitrosamines occurring in processed meat and poultry products. N-nitrosodiethylamine (NDEA), and N-nitrosodibutylamine (NDBA) are also frequently observed to a lesser extent. The processed meat and poultry products with the highest levels of volatile N-nitrosamines were pork (fried, fat only eaten), poultry (fried), poultry (spiced, grilled), and bacon (fried).     

The effect of salt reduction on sensory quality and microbial growth in hotdog sausages,bacon, ham and salami

Sodium chloride (NaCl) is a multi-functional ingredient used to inhibit microbial growth and to ensure good texture and taste in processed meat. This study showed how moderately (22–25%) and greatly (43–50%) reduction of NaCl affected yield, sensory quality and microbial growth in hotdog sausages, bacon, cooked cured ham and salami. In greatly reduced products, the yield was reduced by 8% in sausages and 6% in ham, whereas the yield in bacon and salami remained unaffected. The microbial growth was generally not affected by reducing the content of NaCl to 2.0% in sausages, 2.3% in bacon, 1.7% in ham and 6.3% in salami (aqueous phase). Salt taste, juiciness and texture were the sensory parameters most affected by the NaCl reduction. In sausages and ham, reduction from 2.2% to 1.7% and from 2.3% to 1.3% (w/w), respectively, did not alter the sensory properties. In contrast, the sensory properties of bacon and salami were significantly affected already after a moderately reduction.     

The microbial ecology of a typical Italian salami during its natural fermentation

We have investigated the bacteria and yeast ecology of the typical Italian Ciauscolo salami that is produced in Central Italy using a polyphasic approach based on culture-dependent and -independent methods. The physico-chemical analyses showed a progressive drop in pH and water activity (aw) during ripening. The viable counts revealed a dominance of lactic acid bacteria (LAB) over coagulase negative cocci (CNC) and yeasts. From the molecular identification of the isolates, the prevalence of Lactobacillus curvatus, Lb. plantarum and Staphylococcus xylosus was shown among the bacteria, while Debaryomyces hansenii was the prevalent species among the yeasts, and it was isolated throughout the whole ripening process. Minority species, namely Rhodotorula mucillaginosa and Trichosporon brassicae, were also recovered from the meat batter. The total microbial community was profiled without cultivation by analyzing the DNA that was directly extracted from the salami samples. Moreover, the cultivable community was profiled by analyzing the DNA recovered from bulk cells that were obtained by harvesting the colonies from serial-dilution agar plates. The 16S rRNA gene V1 and V3 regions were used as targets in the denaturing gradient gel electrophoresis (DGGE) profiling of the LAB and CNC communities, respectively, while the diversity and dynamics of the yeast population were assessed by analyzing a portion of the 28S rRNA gene. Our findings suggest that the microbial diversity of fermented meat products can be successfully investigated by this polyphasic approach that is based on the assessment of both the total and the cultivable community diversity.     

Survey of yeasts for antagonistic activity against Salmonella Poona in cantaloupe juice and wounds in rinds coinfected with phytopathogenic molds

Application of yeasts as biocontrol agents to prevent mold decay of fruits and vegetables has been described. We examined 10 yeasts for potential antagonistic activity against survival and growth of Salmonella Poona in cantaloupe juice and decay by Cladosporium cladosporioides and Geotrichum candidum in wounds on cantaloupe rind. Cantaloupe juice was inoculated using five schemes: Salmonella Poona only (1.10 log CFU/ml), high (3.93 to 5.21 log CFU/ml) or low populations (1.79 to 3.26 log CFU/ml) of yeasts only, and Salmonella Poona combined with high or low populations of yeasts. High initial populations of Debaryomyces hansenii, Pichia guilliermondii, and Pseudozyma sp. were antagonistic to Salmonella Poona in cantaloupe juice stored at 20 degrees C for 48 h. Wounds in cantaloupe rinds were inoculated with yeast and mold or yeast, mold, and Salmonella Poona, and cantaloupes were stored at 4 degrees C for 14 days or 20 degrees C for 7 days. The pH of rind tissue inoculated with C. cladosporioides and yeasts increased significantly (P < or = 0.05) at 20 degrees C. Wounds that were inoculated with P. guilliermondii, together with C. cladosporioides or G. candidum, did not show mold growth at 4 and 20 degrees C. Populations of Salmonella Poona (6.40, 7.26, and 7.98 log CFU per sample) were lower in wounds coinoculated with G. candidum and three of the test yeasts (D. hansenii, P. guilliermondii, and Cryptococcus albidus, respectively) compared to coinoculation with G. candidum or the other seven yeasts. Candida oleophila and Rhodotorula glutinis showed the most promise in reducing the population of Salmonella Poona in wounds in rinds of cantaloupes coinoculated with G. candidum and stored at 4 degrees C.     

Identification of fungal contamination and determination of mycotoxigenic molds by micellar electrokinetic capillary chromatography in smoked paprika

The purpose of this work was to analyze the fungal contamination in smoked and unsmoked paprika processed from different cultivars of pepper and to investigate the ability of these and other mycotoxigenic molds to grow and synthesize mycotoxins in smoked paprika. Eighteen mycotoxins were evaluated using micellar electrokinetic capillary chromatography. No relevant differences were found in fungal contamination between smoked and unsmoked paprika. The number of yeasts obtained was low, ranging from 0.4 to 3.29 log CFU g(-1); most of the yeast strains were identified as Cryptococcus spp. followed by Candida spp. All mold counts were <4 log CFU g(-1). Aspergillus, Cladosporium, Penicillium, and Fusarium were the predominant hyphomycete genera. Six mycotoxins were identified in the extracts of several strains isolated from paprika and incubated on malt extract agar. Penicillium expansum followed by Penicillium citrinum and Penicillium raistrickii were the dominant mycotoxigenic fungi isolated. Most of themycotoxin-producing fungi produced detectable amounts of mycotoxins when grown on paprika agar.     

Differentiation of yeasts growing on dry-cured Iberian ham by mitochondrial DNA restriction analysis, RAPD-PCR and their volatile compounds production

The efficiency of mitochondrial DNA (mtDNA) restriction analysis, RAPD-PCR and volatile compounds analysis to differentiate yeast biotypes involved in flavour development of dry-cured Iberian ham throughout the ripening process is evaluated. For this purpose, 86 yeasts isolated from Iberian hams in the main ripening stages at different industries of the four Protected Designations of Origin of this product, were used. The combination of mtDNA restriction analysis and RAPD-PCR using the primer (GACA)₄ showed a higher variability in the yeast species detected than obtained using only mtDNA restriction analysis. Only two species, Debaryomyces hansenii and Candida zeylanoides, were identified throughout the whole ripening process and a wide diversity of biotypes was found in these two species, with those of D. hansenii predominating. Clear differences between biotypes were detected in the generation of volatile compounds, with the biotype C2-2 of D. hansenii showing the highest concentrations of volatiles. The combined use of mtDNA restriction analysis and RAPD-PCR distinguishes yeast biotypes with different production of volatile compounds. In addition, analysis of the production profile of volatile compounds is needed to differentiate yeast strains of the same biotype recovered at different stages of ripening. Thus, the combination of these three methods could be very useful to select or monitor yeasts as starter cultures in dry-cured meat products.     

Survival and growth of Clostridium perfringens in commercial no-nitrate-or-nitrite-added (natural and organic) frankfurters, hams, and bacon

The popularity of "preservative-free" foods among consumers has stimulated rapid growth of processed meats manufactured without sodium nitrite. The objective of this study was to quantify the potential for Clostridium perfringens growth in commercially available processed meats manufactured without the direct addition of nitrite or nitrate. Commercial brands of naturally cured, no-nitrate-or-nitrite-added frankfurters (10 samples), hams (7 samples), and bacon (9 samples) were obtained from retail stores and challenged with a three-strain inoculation (5 log CFU/g) of C. perfringens. Reduced inhibition (P < 0.05) was observed in seven brands of frankfurters, six brands of hams, and four brands of bacon when compared with each respective sodium nitrite-added control. In naturally cured and truly uncured commercial frankfurters, growth over time was approximately 4.7 log, while conventionally cured frankfurters exhibited growth at 1.7 log. Naturally cured ham and bacon products exhibited growth at 4.8 and 3.4 log, respectively, while their conventionally cured counterparts exhibited growth at 2.6 and 2.3 log, respectively. These products also demonstrated variation in growth response. The results indicate that commercially available natural/organic naturally cured meats have more potential for growth of this pathogen than do conventionally cured products. Natural and organic processed meats may require additional protective measures in order to consistently provide the level of safety from bacterial pathogens achieved by conventionally cured meat products, and which is expected by consumers.     

金华火腿发酵过程中微生物区系研究

本实验对金华火腿传统工艺发酵过程中微生物的数量及种类的变化进行了较详细深入的研究,检测了一年中金华火腿表面和深部微生物的数量变化规律、微生物的种类,并检测了发酵过程中水分、含盐量和pH值的动态变化.研究结果表明:金华火腿前期内部及表面微生物数量极少,晾晒时表面微生物数量增加,进入发酵室后内部微生物也很快增加达到106CFU/g,火腿成熟生香时期,内部微生物数量降至103CFU/g以下,优势菌相是葡萄球菌,其次是乳酸菌;内部酵母菌群中占优势的菌种主要有类筒假丝酵母、汉逊德巴利酵母、赛道威汉逊酵母、红酵母等,火腿表面的霉菌数量影响着内部的微生物.最初的阶段,二者一起增加,当霉菌的数量增加到一定水平时,抑制内部微生物的生长,使其数量减少;当霉菌数量减少时,内部微生物数量就会增加,发酵前期青霉菌占优势,主要有意大利青霉、简单青霉、灰绿青霉、柑桔青霉等.发酵后期,为曲霉菌占优势,主要包括萨氏曲霉、灰绿曲霉、黄柄曲霉等.金华火腿独特的风味与该地区特有的微生物区系有一定关系.     

Occurrence and characterization of yeasts isolated from artisanal Fiore Sardo cheese

The occurrence of yeast microflora in artisanal Fiore Sardo cheese during ripening was studied. Mean yeast counts ranged from 2.64+/-1 log(10) cfu ml(-1) in milk to 0.65+/-1 log(10) cfu g(-1) in 9 months cheese, with the higher counts observed in 48-h-old cheese. Strains belonging to the prevalent species Debaryomyces hansenii, Kluyveromyces lactis, Geotrichum candidum, Candida zeylanoides and Candida lambica were selected for technological and genotypic characterization. All D. hansenii strains fermented glucose and assimilated lactate, a high percentage assimilated citrate and only a few showed proteolytic and lipolytic activity. All K. lactis strains were able to both assimilate and ferment lactose, to assimilate lactate and to exhibit proteolytic activity on casein. G. candidum assimilated lactate and some strains showed proteolytic and lipolytic activity. C. zeylanoides showed lipolytic activity on tweens and the majority of strains assimilated citrate. C. lambica fermented glucose and assimilated lactate. Considering their diffusion and technological characteristics, an important role for K. lactis and G. candidum in the early stages of the ripening process and for D. hansenii after the first month of ripening can be suggested. RAPD-PCR analysis with M13 primer grouped the isolates in well-separated clusters with their type strains and confirmed the previous phenotypic identification. The high intraspecific homogeneity observed in tested strains could be explained by their isolation from a common substrate and from neighbouring geographical areas. This preliminary study allowed us to isolate autochthon yeast strains showing particular properties which can contribute to the production of typical cheese taste and flavour.     

Dynamics and diversity of non-Saccharomyces yeasts during the early stages in winemaking

This detailed study observed the yeasts present in the ecological niche of "wine must". The dynamics and identity of non-Saccharomyces yeasts during the cold maceration and alcoholic fermentation of grape must were investigated under real production conditions in the Bordeaux region. Furthermore, we studied the impact of two oenological parameters on the development and diversity of non-Saccharomyces yeasts during cold maceration: temperature management and the timing of dried yeast addition. The non-Saccharomyces community underwent constant changes throughout cold maceration and alcoholic fermentation. The highly diverse non-Saccharomyces microflora was present at 10(4)-10(5) CFU/mL during cold maceration. The population increased to a maximum of 10(6)-10(7) CFU/mL at the beginning of alcoholic fermentation, then declined again at the end. The population at this point, evaluated at around 10(3)-10(4) CFU/mL, was shown to be dependent on the timing of yeast inoculation. The choice of temperature was the key factor for controlling the total yeast population growth, as well as the species present at the end of cold maceration. Hanseniaspora uvarum was a major species present in 2005 and 2006, while Candida zemplinina was very abundant in 2006. A total of 19 species were isolated.     

Characterisation of yeast flora isolated from an artisanal Portuguese ewes' cheese

The evolution of the yeast flora was studied for an artisanal semi-hard ewes' cheese made from raw milk. Mean log10 yeast counts per gram of cheese body ranged from 2.7 to 6.4, with the higher counts observed after a ripening period of 30 days. The yeast population decreased thereafter and, at the end of curing process, reached values similar to those of the beginning. A total of 344 yeasts strains were randomly isolated from the curd and cheese body during the 60 days long ripening period. Esterase activity was common to almost all isolates (98%) while proteolysis was observed in 12% of the total yeast population. The proportion of strains with positive glucose fermentation increased from 21% in the curd to 75% at the end of the ripening period. A total of 150 isolates representative of the physiological characteristics tested were examined with the API ID 32C system showing different degrees of quality of identification. Only 15% of the strains (23 isolates) were excellently identified being assigned to the species Candida zeylanoides. The most frequent species appeared to be Debaryomyces hansenii (anamorph Candida famata) and Candida intermedia. These two species amounted to 9% of the yeasts in the curd increasing to 86% at the end of the ripening period.     

Application of enterocins as biopreservatives against Listeria innocua in meat products

The antilisterial effect of enterocins A and B in meat and meat products (cooked ham, minced pork meat, deboned chicken breasts, paté, and slightly fermented sausages [espetec]) have been shown. An infective dose of 5 to 10 most probable numbers (MPN)/g to simulate the counts of Listeria generally found in meat products was used. Enterocins at 4,800 AU/g reduced the numbers of Listeria innocua by 7.98 log cycles in cooked ham and by 9 log cycles in paté when stored at 7 degrees C for 37 days. In deboned chicken breasts stored at 70 degrees C for 7 days, 4,800 AU/cm2 of enterocins diminished the L. innocua counts in 5.26 log cycles when compared to the control batch. In minced pork meat held at 7 degrees C for up to 6 days, 1,600 AU/g kept L. innocua counts under 3 MPN/g, while the control batch reached 50 CFU/g. In espetec sausages, 648 AU/g diminished the number of L. innocua under 50 CFU/g from the fifth day until the end of the process (12 days) while the control batch kept the initial counts (3 x 104 CFU/g). This is the first report on enterocins showing an antilisterial effect in different types of meat products.     

Prevalence and concentration of Listeria monocytogenes in sliced ready-to-eat meat products in the Hellenic retail market

The aim of this work was to estimate the prevalence and concentration of Listeria monocytogenes in packaged precut (slices or cubes) ready-to-eat (RTE) meat products available in the Hellenic retail market. Samples of these RTE meat products (n = 209) were taken from local supermarkets during a 3-month period and analyzed for the presence of L. monocytogenes with an automated enzymatic qualitative immunoassay followed by biochemical confirmation of positive results. The concentration of the pathogen in the positive samples was also determined. Seventeen samples (8.1%) were positive for L. monocytogenes. Eight (47.1%) of these 17 samples were from the same manufacturer; 36.4% of the products tested from this manufacturer were positive for L. monocytogenes. When bacon samples were not considered, the estimated prevalence of L. monocytogenes in sliced RTE meat products was much lower (3.1%). The L. monocytogenes populations in all positive samples were low, < or = 10 CFU/g. In 64.7% of the L. monocytogenes-positive samples, other Listeria species, including L. innocua and L. welshimeri, were also present at <10 to 690 CFU/g. These results indicate that L. monocytogenes is present in low numbers but is in a considerable proportion of the packaged precut RTE meat products that are sold in the Hellenic retail market. Cooked ham and bacon cut in cubes were the sample types most often contaminated with L. monocytogenes. The higher level of handling (e.g., cutting) associated with these products may further increase the risk of contamination with L. monocytogenes.     

Effect of Penicillium chrysogenum and Debaryomyces hansenii on the volatile compounds during controlled ripening of pork loins

During ripening of meat products such as dry-cured ham, the moulds and yeasts that proliferate on the surface may contribute to flavour development. However, their contribution to volatile components of dry-cured meat products is not known. One strain each of Penicillium chrysogenum and Debaryomyces hansenii, selected from dry-cured ham by their proteolytic activity, were tested to determine their effect on the volatile compounds during ripening. Sterile pork loins were inoculated and ripened for 106 days. Volatile compounds collected with a Solid Phase Micro-Extraction (SPME) fibre were analysed by GC/MS. Inoculation of pork loins with P. chrysogenum lead to a decrease in compounds attributed to lipid oxidation and to an increase of compounds derived from free amino acids. Inoculation with D. hansenii seemed to favour the formation of complex alcohols.     

Survival of dairy-associated yeasts in yoghurt and yoghurt-related products

The poor survival of probiotic bacteria added to yoghurts is mainly attributed to the low pH of the product environment. Since yeasts have the ability to metabolize organic acids, resulting in a decrease in acidity, the inclusion of yeasts as part of the normal microflora, in association with probiotic bacteria has been suggested with the intention to assure better survival of the probiotic organisms in bio-yoghurt. Furthermore, a commensalistic association between yeasts and lactic acid bacteria exists. In order to understand the potential impact of yeast on probiotic bacteria, it was firstly important to assess the ability of yeast isolates to grow and survive in yoghurt.Accordingly, four dairy-associated yeasts, Debaryomyces hansenii, Kluyveromyces marxianus, Yarrowia lipolytica and Issatchenkia orientalis, associated commonly with yoghurt were isolated and inoculated subsequently into yoghurt and related dairy products during processing. The survival and growth of the yeasts were monitored over a 4-week storage period, the normal time accepted as the shelf-life of yoghurts. pH, sugar utilization and the production of organic acids were determined on a regular basis during the shelf-life to evaluate the possible contribution of the yeasts towards the products. The yeast species were able to survive in bio-yoghurt reaching maximum counts exceeding 10⁷ cfu g⁻¹. Despite the inability of some species to utilize lactose, the yeast species utilized available organic acids, galactose and glucose derived from bacterial metabolism of the milk lactose, as well as possible free fatty acids or free amino acids present in the dairy products. Excessive gas and ethanol production initiated by some yeast species proved, however, to be major constraints.     

Identification of yeasts associated with milk products using traditional and molecular techniques

An integrated approach including phenotypic (morphological, biochemical and physiological characterization) and genotypic (RAPD-PCR, sequencing of D1/D2 domain of 26S rRNA encoding gene) methods was used for the identification of yeasts isolated from different milk products. There were 513 isolates in all, 460 ascomycetous and 53 basidiomycetous yeasts. The yeast isolates were characterized on the basis of their biochemical and physiological properties, and the D1/D2 domain of 26S rDNA was sequenced in selected strains. Relying on the obtained results from both the data-sets, corresponding type strains were selected and compared with the respective yeast isolates from milk products by RAPD fingerprinting. The strains showing a degree of similarity >80% were considered conspecific. By means of the applied techniques it was possible to identify 92% yeast isolates at species level. Debaryomyces hansenii, Geotrichum candidum, Kluyveromyces marxianus, Yarrowia lipolytica and Candida zeylanoides are the most frequently isolated species. The majority of the yeasts were isolated from fresh and sour curd cheese. A comparison of the results obtained by phenotypic and genotypic investigation revealed that the identification based on classical methods was supported by genotypic characterization in only 54% of examined isolates. The results described in this work show that the applied molecular identification is a reliable approach to the identification of yeasts associated with milk products in contrast to the conventional biochemical and physiological tests. The identification of new yeast species requires additional genetic markers such as sequencing of different genes or DNA:DNA hybridization.     

内蒙古和新疆牧区酸马奶中酵母菌的多样性及其优势菌发酵特性

探究内蒙古和新疆牧区传统酸马奶中酵母菌的多样性及其优势菌的发酵特性,依据GB 4789.15-2010《食品微生物学检验 霉菌和酵母计数》进行菌株分离,再应用26S rDNA D1/D2区序列和内转录间隔区（internaltranscribed space,ITS）序列分析技术鉴定分离株,并依据产酒精能力、产酸能力、风味及质构特性等指标筛选优良菌株。结果表明：1）内蒙古酸马奶样品中酵母菌活菌数为105 CFU/mL,新疆样品在105～107 CFU/mL之间。两地样品中共鉴定出包括Kluyveromyces marxianus、Pichia fermentans、Pichia cactophila、Candida zeylanoides在内的酵母菌13 种。两地间样品酵母菌组成不尽相同,具有地区差异性,内蒙古优势菌为Kluyveromyces marxianus、Pichiacactophila和Pichia fermentans,新疆优势菌为Candida zeylanoides和Kluyveromyces marxianus。2）优选出的4 株酵母菌有2 株为Pichia cactophila,另外2 株为Kluyveromyces marxianus和Candida zeylanoides,发酵特性良好,可用于新型发酵乳制品开发。