Experimental effect of ozone upon the microbial flora of commercially produced dairy fermented products.

Ozone was used to control spoilage microorganisms during the manufacturing of dairy products. Ozone stream was applied onto the surface of freshly filled yoghurt cups just before storage for curd development in order to prevent cross contamination from spoilage airborne microorganisms. Accordingly, brine solution was bubbled with ozone for various periods of time and used for ripening of white (feta type) cheese. Both products were subjected to a continuous monitoring of microbial load and also tested for their sensorial properties. In ozonated yoghurt samples there was a reduction in mould counts of approximately 0.6Logcfu/g (25.1%) by the end of the monitoring period in relation to the control samples. In white cheese ripened with ozonated brine (1.3mg/L O3, NaCl 5%) it seems that ozone treatment during the two months of observation reduced some of the mould load but without offering any advantages over the use of traditional brine (NaCl 7%). However, some sensorial alterations were observed, probably due to the organic load in the brine which deactivates ozone in early stages of application. It is concluded that, if the factors of time and concentration of ozone are configured properly, ozonation could be a promising approach safeguarding the production of some dairy products.

Efficacy of natamycin to control fungal growth in natural black olive fermentation.

The effect of natamycin as a fungal control agent in natural black olive fermentation according to the traditional anaerobic system was studied. Black Conservolea olives were subjected to spontaneous fermentation in 8% (w/v) NaCl brine solution (control treatment) or brine supplemented with 0.01% (w/v) natamycin (active compound) for an overall period of 60 days. The changes in the microbial association (lactic acid bacteria, yeasts, Enterobacteriaceae), pH, titratable acidity, organic acids, and volatile compounds were monitored throughout fermentation. The initial microbiota consisted of lactic acid bacteria, yeasts, and Enterobacteriaceae. The addition of natamycin in the brine inhibited the growth of yeasts, without affecting the population dynamics of bacteria, resulting in a more vigorous fermentation with higher titratable acidity compared to spontaneous control process. Moreover, the presence of natamycin inhibited mould spoilage caused by the development of fungal mycelium on the surface of the brine during the traditional anaerobic fermentation system employed widely in Greece in natural black olive processing. Natamycin could be an important component of a processing system to control fungal growth in natural black olive fermentation and at the same time it has the potential to enhance the process by favouring the growth of the indigenous population of lactic acid bacteria against other competing microorganisms.

Thermally-dried immobilized kefir on casein as starter culture in dried whey cheese production.

The aim of the present study was to evaluate the use of thermally-dried immobilized kefir on casein as a starter culture for protein-enriched dried whey cheese. For comparison reasons, dried whey cheese with thermally-dried free kefir culture and with no starter culture were also produced. The effect of the nature of the culture, the ripening temperature and the ripening process on quality characteristics of the whey cheese was studied. The association of microbial groups during cheese maturation suggested repression of spoilage and protection from pathogens due to the thermally-dried kefir, as counts of coliforms, enterobacteria and staphylococci were significantly reduced in cheeses produced using thermally-dried kefir starter cultures. The effect of the starter culture on production of volatile compounds responsible for cheese flavor was also studied using the SPME GC/MS technique. Thermally-dried immobilized kefir starter culture resulted in an improved profile of aroma-related compounds. The preliminary sensory evaluation ascertained the soft, fine taste and the overall improved quality of cheese produced with the thermally-dried immobilized kefir. The potential of protein-based thermally-dried starter cultures in dairy products is finally highlighted and assessed.

Whey-cheese production using freeze-dried kefir culture as a starter.

AIMS: The aim of the present study was to evaluate the use of a freeze-dried kefir culture in the production of a novel type of whey-cheese similar to traditional Greek Myzithra-cheese, to achieve improvement of the quality characteristics of the final product and the extension of shelf-life. METHODS AND RESULTS: The use of kefir culture as a starter led to increased lactic acid concentrations and decreased pH values in the final product compared with whey-cheese without starter culture. The effect of the starter culture on production of aroma-related compounds responsible for cheese flavour was also studied using the solid phase microextraction gas chromatography/mass spectrometry technique. Spoilage in unsalted kefir-whey-cheese was observed on the thirteenth and the twentieth day of preservation at 10 and 5 degrees C, respectively, while the corresponding times for unsalted whey-cheese preservation were 11 and 14 days. CONCLUSIONS: The cheeses produced were characterized as high-quality products during the preliminary sensory evaluation. An indication of increased preservation time was attributed to the freeze-dried kefir culture, which also seemed to suppress growth of pathogens. SIGNIFICANCE AND IMPACT OF THE STUDY: The results suggested the use of kefir culture as a means to extend the shelf-life of dairy products with reduced or no salt content.

An economic evaluation of freeze-dried kefir starter culture production using whey.

An economic study is presented in which industrial-scale production of freeze-dried kefir starter culture is discussed based on results on a laboratory scale. Industrial scale-up was based on a 3-step process using 3 bioreactors of 100, 3,000, and 30,000 L for 300 kg of freeze-dried culture/d of plant capacity. The major cost component of the total investment was the freeze-drying machinery, which consisted of 57% of the total investment. Production cost was reduced from 15.4 euros/kg ($18.5/kg) to 2.9 euros/kg ($3.5/kg) when the production capacity was increased from 30 to 900 kg/d. An economic analysis revealed a 3.5-fold increase in production cost compared with the corresponding production cost of the wet product, with an added value of up to 10.8 x 10(9) euros ($13.0 x 10(9)) within the European Union.

Evaluation of freeze-dried kefir coculture as starter in feta-type cheese production.

The use of freeze-dried kefir coculture as a starter in the production of feta-type cheese was investigated. Maturation of the produced cheese at 4 degrees C was monitored for up to 70 days, and the effects of the starter culture, the salting method, and the ripening process on quality characteristics were studied. The use of kefir coculture as a starter led to increased lactic acid concentrations and decreased pH values in the final product associated with significantly higher conversion rates compared to salted rennet cheese. Determination of bacterial diversity at the end of the ripening process in salted kefir and rennet cheeses by denaturing gradient gel electrophoresis technology, based on both DNA and RNA analyses, suggested a potential species-specific inhibition of members of the genera Staphylococcus and Psychrobacter by kefir coculture. The main active microbial associations in salted kefir cheese appeared to be members of the genera Pseudomonas and Lactococcus, while in salted rennet cheese, Oxalobacteraceae, Janthinobacterium, Psychrobacter, and Pseudomonas species were noted. The effect of the starter culture on the production of aroma-related compounds responsible for cheese flavor was also studied by the solid-phase microextraction-gas chromatography-mass spectrometry technique. Kefir coculture also appeared to extend the shelf life of unsalted cheese. Spoilage of kefir cheese was observed on the 9th and 20th days of preservation at 10 and 5 degrees C, respectively, while spoilage in the corresponding rennet cheese was detected on the 7th and 16th days. Microbial counts during preservation of both types of unsalted cheese increased steadily and reached similar levels, with the exception of staphylococci, which were significantly lower in unsalted kefir cheese. All types of cheese produced with kefir as a starter were approved and accepted by the panel during the preliminary sensory evaluation compared to commercial feta-type cheese.

Probiotic cheese production using Lactobacillus casei cells immobilized on fruit pieces.

Lactobacillus casei cells were immobilized on fruit (apple and pear) pieces and the immobilized biocatalysts were used separately as adjuncts in probiotic cheese making. In parallel, cheese with free L. casei cells and cheese only from renneted milk were prepared. The produced cheeses were ripened at 4 to 6 degrees C and the effect of salting and ripening time on lactose, lactic acid, ethanol concentration, pH, and lactic acid bacteria viable counts were investigated. Fat, protein, and moisture contents were in the range of usual levels of commercial cheeses. Reactivation in whey of L. casei cells immobilized on fruit pieces after 7 mo of ripening showed a higher rate of pH decrease and lower final pH value compared with reactivation of samples withdrawn from the remaining mass of the cheese without fruit pieces, from cheese with free L. casei, and rennet cheese. Preliminary sensory evaluation revealed the fruity taste of the cheeses containing immobilized L. casei cells on fruit pieces. Commercial Feta cheese was characterized by a more sour taste, whereas no significant differences concerning cheese flavor were reported by the panel between cheese containing free L. casei and rennet cheese. Salted cheeses scored similar values to commercial Feta cheese, whereas unsalted cheese scores were significantly lower, but still acceptable to the sensory panelists.

High-temperature wine making using the thermotolerant yeast strain Kluyveromyces marxianus IMB3.

Kluyveromyces marxianus IMB3 yeast cells were immobilized on delignified cellulosic material, apple, and quince separately. Both immobilized and free cells were used in high-temperature wine making, and their fermented grape must contained 3 to 4% alcohol. Semisweet wines were produced by the addition of potable alcohol to the fermented must. Preliminary sensory evaluation of the produced semisweet wines showed good flavor and aroma. The final product contained extremely low levels of higher and amyl alcohols while ethyl acetate was at levels usually present in wines. The ferment produced may be blended with other products to improve their quality.

Enhanced bioremediation of n-alkane in petroleum sludge using bacterial consortium amended with rhamnolipid and micronutrients.

The purpose of the present study was to investigate possible methods to enhance the rate of biodegradation of oil sludge from crude oil tank bottom, thus reducing the time usually required for bioremediation. Enhancement of biodegradation was achieved through bioaugmentation and biostimulation. About 10% and 20% sludge contaminated sterile and non-sterile soil samples were treated with bacterial consortium (BC), rhamnolipid biosurfactant (RL) and nitrogen, phosphorus and potassium (NPK) solution. Maximum n-alkane degradation occurred in the 10% sludge contaminated soil samples. The effects of treatment carried out with the non-sterile soil samples were more pronounced than in the sterile soils. Maximum degradation was achieved after the 56th day of treatment. n-Alkanes in the range of nC8-nC11 were degraded completely followed by nC12-nC21, nC22-nC31 and nC32-nC40 with percentage degradations of 100%, 83-98%, 80-85% and 57-73% respectively. Statistical analysis using analysis of variance and Duncan's multiple range test revealed that the level of amendments, incubation time and combination of amendments significantly influenced bacterial growth, protein concentration and surface tension at a 1% probability level. All tested additives BC, NPK and RL had significant positive effects on the bioremediation of n-alkane in petroleum sludge.

High-temperature alcoholic fermentation of whey using Kluyveromyces marxianus IMB3 yeast immobilized on delignified cellulosic material.

A novel system for high-temperature alcoholic fermentation of whey is described. This system consists of Kluyveromyces marxianus yeast immobilized on delignified cellulosic material (DCM). The effect of pH, initial lactose concentration and temperature on the fermentation of a synthetic medium containing lactose was studied. Batch fermentations of whey were also carried out and the formation of volatile by-products was examined. The concentrations of higher alcohols were found to be in very low levels leading to a product of improved quality. The fermented whey had an improved characteristic aroma compared to unfermented whey. The possibility to use fermented whey as raw material for the production of a novel, low alcohol content drink was also investigated.

Continuous whey fermentation using kefir yeast immobilized on delignified cellulosic material.

Delignified cellulosic-supported biocatalyst, prepared by immobilization of kefir yeast on delignified cellulosic material (DCM), was found to be suitable for continuous, modified whey fermentation. The modified whey contained 1% raisin extract and molasses. Ethanol productivities ranged from 3.6 to 8.3 g L(-1)day(-1), whereas parameters such as ethanol concentration, residual sugars, and daily fermented whey productivity were acceptable for the production of potable alcohol and alcoholic drinks in industrial fermentations. The continuous fermentation bioreactor was operated for 39 days, stored for 18 days at 4 degrees C, and operated again for another 15 days without any diminution of the ethanol productivity. The concentrations of higher alcohols (propanol-1, isobutyl alcohol, and amyl alcohols) were low. The main volatile byproducts formed in the continuous process were similar to those observed in alcoholic beverages, and the fermented whey had a good aroma. The concentrations of higher alcohols were very low when compared to that of ethyl acetate, therefore resulting in a quality product. The possibility of using such a process for the production of potable alcohol or a novel, low-alcohol content drink is proposed.

Wine production using yeast immobilized on apple pieces at low and room temperatures.

A biocatalyst was prepared by immobilization of Saccharomyces cerevisiae strain AXAZ-1 on apple pieces. It was examined by electron microscope and studied during the fermentation of grape must for batch wine-making. The immobilized yeast showed an important operational stability without any decrease of its activity even at low temperatures (1-12 degrees C). Specifically, at 6 degrees C the biocatalyst favored wine production within 8 days, which is less time than is required for the natural fermentation of grape must. At 1 degrees C wine production was effected in 1 month. This speeding up of the fermentation could be accepted and adopted by the industry for scaling up the wine-making process. Total and volatile acidities were similar to those found in dry wines. The concentrations of higher alcohols (propanol-1 and isobutyl alcohol) were low. The presence of amyl alcohols proved to be temperature dependent and decreased with the temperature decrease. The values of ethyl acetate concentrations were relatively high, up to 154 mg/L. This probably contributes to the fruity aroma and excellent taste of the produced wines. There was no indication of vinegar odor in the product given that the volatile acidity was <0.47 g of acetic acid/L. From the GC-MS analysis it was concluded that cell immobilization did not create serious changes in the product (wine) with regard to the qualitative composition of the aroma compounds.