Behaviour of Listeria monocytogenes inoculated into Minas Frescal cheese made by direct acidification or lactic culture during refrigerated storage

The behaviour of Listeria monocytogenes Scott A artificially inoculated into Brazilian Minas Frescal cheese manufactured with the addition of a lactic culture or by direct acidification with lactic acid was evaluated at 5 and 10°C for 25 days. In cheese produced with a lactic culture, the counts remained almost unchanged during storage, whereas cheeses produced by direct acidification presented an increase in the counts of 2–3 log cycles. The profiles of the lactic bacterial counts, associated with those of L. monocytogenes, show a strong influence of the competitive starter culture on the survival of L. monocytogenes . This suggests that the addition of starter culture is an efficient way to control the growth of L. monocytogenes in these products.     

USE OF MICROPOROUS PLASTIC FOR IN-PACKAGE CONTROLLED ATMOSPHERES

Ground beef packaged and stored with carbon dioxide-generating reagent packets made from (1) microporous plastic film or (2) non-woven cellulose cloth had significantly (p<0.01) lower bacterial counts than ground meat packaged and stored without packets. There was no significant difference (p>0.05) in bacterial counts of samples stored with the 2 types of packets. Samples with in-package generated carbon dioxide atmospheres had approximately 2–3 days additional shelf-life before spoilage.     

Influence of the frozen storage on some characteristics of ripened Manchego-type cheese manufactured with a powdered vegetable coagulant and rennet

A study was made of the effect of freezing and nine months’ frozen storage on the chemical, microbiological and sensorial characteristics of fully ripened Manchego-type cheese. Chemical components, water activity and nitrogen compounds were not altered by freezing and frozen storage. Soluble and non-protein nitrogen were significantly higher in cheeses produced with the powdered vegetable coagulant (PVC) compared to those made with rennet. Proteolysis continued slowly during frozen storage, with higher amino acid and ammonia nitrogen rates at the end of the storage period. Micrococci (P-value <0.05, in cheese made with PVC) and staphylococci counts tended to decrease during frozen storage; however total viable, lactic acid bacteria and mold-yeasts counts were similar throughout the nine months. Changes in sensorial parameters were observed (P-value <0.05) throughout freezing and frozen storage, affecting the creaminess of cheeses made with chymosin and the taste intensity and salty taste of cheese obtained with vegetable coagulant. Cheeses obtained with PVC proved to have a more acid taste and greater taste intensity (P < 0.01) as well as displaying a more bitter taste, greater creaminess and lower degree of hardness (P-value <0.001) than those produced with rennet.     

Chemical composition and microbial evaluation of Argentinean Corrientes cheese

The chemical and microbial composition of an artisanal cheese made from raw cow's milk produced and consumed in the province of Corrientes (north-eastern Argentina) was evaluated using standard methods. Corrientes cheese has high moisture content (50–60%), normal protein and fat contents (21–27 and 22–26% respectively), and is low in salt (0.5–2.0% w/w). Microbial counts also varied significantly between samples (colony-forming units per gram ranges covering logs of 5–11), probably due to environmental contamination in the raw material. These results will help produce higher quality Corrientes cheeses with well-defined characteristics.     

The use of halloum cheese for the production of processed cheese

Halloum is a white, semi-hard cheese made in a number of countries around the Mediterranean and in the Middle East from raw milk. In this study, halloum was made from natural raw milk and from milk acidified to pH 4.5 before renneting. Processed cheese was made from the two types of halloum using 4.2% emulsifying salts and 20–25% added water. There was an increase in protein, fat and total solids in the processed cheese made from acidified halloum, but a taste panel gave a higher sensory rating to the processed cheese based upon normal halloum.     

Evaluation of microbial survival post-incidence on fresh Mozzarella cheese

Commercial fresh Mozzarella cheese is made by direct acidification and is stored dry or in water without salt addition. The cheese has a shelf life of 6 wk, but usually develops an off-flavor and loses textural integrity by 4 wk, potentially due to the lack of salt and high moisture that allow the outgrowth of undesirable bacteria. To understand how microbial incidence affects cheese quality and how incident pathogen-related bacteria are limited by salt level during refrigerated storage, we made fresh Mozzarella cheese with high (2%) and low (0.5%) salt. The high-salt cheese was packaged and stored dry. The low-salt cheese was packaged and stored either dry or in 0.5% salt brine. One portion of cheeses was evaluated for surviving incident microbes by aerobic plate counts, coliform counts, and psychrophilic bacterial counts, of which coliforms and psychrophiles were not detected over 9 wk. Aerobic plate counts remained at 100 to 300 cfu/g up to 2 wk but increased by 1,000- to 10,000-fold between 4 and 6 wk at all salt levels and storage conditions. Other portions of cheeses were inoculated with either Escherichia coli or Enterococcus faecalis, both of which increased by 100-fold over 90 d of storage. Interestingly, E. coli added to the cheese brine first grew in the brine by 100-fold before attaching to the cheese, whereas Ent. faecalis attached to the cheese within 24 h and grew only on the cheese. We conclude that incident bacteria, even from similar environments, may attach to cheese curd and survive differently in fresh Mozzarella cheese than in brine. Overall, 2% salt was insufficient to control bacterial growth, and slow-growing, cold- and salt-tolerant bacteria may survive and spoil fresh Mozzarella 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°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.     

Bacteriological Aspects of Microfiltration of Cheese Whey,

Microfiltration of cheese whey using 1.2-μm pore size membranes reduced bacterial counts by one to three times. Increased fat concentration in the feed stream governed the decrease in bacterial counts. Fat was trapped onto the membrane proportionally to its amount in the feed stream, and thus, formed a barrier to microorganism penetration into the permeate.     

Aroma–taste interactions between a model cheese aroma and five basic tastes in solution

The flavour perception of cheese results from complex sensory interactions between tastes and aromas. Using a model cheese solution, this study investigated perceived interactions between each of five basic tastes and a cheese aroma mixture containing ten volatile compounds commonly found in cheese. The five tastes – sucrose (sweetness), sodium chloride (NaCl) (saltiness), monosodium glutamate (MSG) (umami), lactic acid (sourness), and caffeine (bitterness) – were individually mixed with cheese aroma in water using a 5 taste level (0.2 log series) by 3 aroma level (0.5 log series) design. Aroma controls with no added taste were also included. This resulted in 18 samples for each single taste–aroma combination. An additional 18 samples were produced using a mixture of all 5 tastes with the 3 aroma levels. A panel of trained assessors (n = 10) evaluated cheese flavour intensity and taste intensity using 100 point line scales. Evaluation was carried out in duplicate, with samples grouped by taste type; 1 evaluation session per taste per replicate. Within type, order of presentation was balanced, and taste type order was randomised between replicates. Cheese flavour intensity was enhanced by sucrose and NaCl, while being suppressed by lactic acid. NaCl enhanced cheese flavour intensity the most at high aroma level, while lactic acid suppressed the most at low aroma level. When MSG level was increased, cheese flavour intensity was enhanced at both low and medium aroma levels, but was suppressed at the high aroma level. The greatest enhancement of cheese flavour intensity was found with the mixture of 5 tastes. Aroma significantly enhanced umami and bitterness, but did not enhance sweetness, saltiness, or sourness. This study showed that the perceived interaction between taste and cheese aroma depended on taste type and on the concentration levels of both taste type and aroma. The mixture of tastes was more effective at enhancing cheese flavour intensity than single tastes. This study provides knowledge that will underpin further study of taste–aroma interactions in a model cheese that aims to optimise cheese flavour intensity and character.     

Role of the microbial population on the flavor of the soft-bodied cheese Torta del Casar

The purpose of this work was to investigate the influence of the spontaneous microbial population on the flavor of Torta del Casar cheese. A total of 16 batches of cheeses with different microbial qualities were used. Their physicochemical and microbial characteristics were evaluated during ripening and then related with the volatile compounds, taste, and flavor properties of the finished cheeses. Acids were the most abundant volatile compounds, followed by alcohols and carbonyls. The amount of acetic acid and several alcohols were linked to cheeses with higher counts of lactic acid bacteria (LAB), whereas Enterobacteriaceae counts were associated with semivolatile fatty acids. The gram-positive catalase-positive cocci counts were correlated with esters and methyl ketones. Although the role of the LAB in the flavor development of Torta del Casar is the most relevant, other microbial groups are necessary to impart the flavor of the cheese and to minimize the possible off-flavor derived from excessive concentrations of LAB metabolites, such as acetic acid.     

The manufacture of Ras cheese from gamma irradiated milk

Cheese milk exposed to gamma irradiation at a dose of 0·5 Mrad was used in the manufacture of Ras cheese. Quality and ripening of cheese made from irradiated milk were compared with those of cheese made from heat-treated milk.Gamma irradiation of cheese milk reduced the total bacterial count and spore formers by 98·98% and 95·77%, respectively, and eliminated the coliform and pathogenic bacteria. This treatment slightly increased the acid and peroxide values of milk fat, as well as the time required for complete coagulation of milk.An oxidized flavour in the fresh and 1-month-old gamma-irradiated milk (GIM) cheese disappeared during ripening. At the end of ripening, GIM cheese had a better consistency and increased flavour compared with cheese made from heated milk.Ras cheese made from GIM differed little in gross chemical composition but contained higher concentrations of soluble nitrogen compounds and of Free Fatty Acids than the control. Irradiation had a stimulating effect on the total, proteolytic and lipolytic bacterial counts during cheese ripening.     

Effects of Lactobacillus strains on the ripening and organoleptic characteristics of Arzúa-Ulloa cheese

Seven batches of Arzúa-Ulloa, a short-ripened soft cow's milk cheese produced in Galicia (NW Spain), were prepared from pasteurized milk. Two control batches of cheese (CB) were made with an acid-aromatic starter containing Lactococcus lactis subsp. lactis and Lactococcus lactis subsp. lactis var. diacetylactis, isolated from raw-milk Arzúa-Ulloa cheeses. Five batches of cheese (LB) were made with the acid-aromatic starter plus one of five strains of mesophilic homofermentative Lactobacillus spp.: four of them isolated from raw-milk Arzúa-Ulloa cheese (characterized in previous works) and the remaining was a commercial Lactobacillus strain. Higher counts of mesophilic viable bacteria, lactic acid bacteria and citrate-fermenting bacteria were found on days 1 or 15 of ripening, while higher counts of lactobacilli were found on day 30 of ripening. On day 1 of ripening the highest diacetyl-acetoin content was noted in the CB, but after day 15 the diacetyl-acetoin content was similar or higher in three of the five LB. The mean degradation of beta-casein in CB was higher than in LB, while the degradation of alpha(s1)-casein was higher in LB. The mean contents of nitrogen-soluble fractions were slightly higher in the LB than in the CB. Volatile free fatty acid (VFFA) contents were, in general, greater in LB than in CB and maximum amounts were determined on day 15 of maturation. Sensorial analysis indicated a more acid taste was in LB, while bitter and astringent tastes were more intense in CB. A positive correlation was found between beta-casein degradation and bitter taste. Yogurt and butter aromas were more intense in CB and in two of the five LB. Firmness was lower in LB and a negative correlation was found between this parameter and alpha(s1)-casein degradation. Crumbliness showed a positive correlation with beta-casein degradation. The use of the Lactobacillus strains assessed in this study is recommended for Arzúa-Ulloa cheese manufacture, in order to enhance the desirable characteristics of this cheese variety, i.e., a soft texture due to alpha(s1)-casein proteolysis but without the bitter taste due to beta-casein degradation and a spicy and slightly rancid aroma and taste.     

Cheddar cheese taste can be reconstructed in solution using basic tastes

The taste of cheese contributes to flavour character directly and by cross-modal interactions with aroma. However, the relative contribution of specific tastes, i.e., sweet, salt, umami, sour, and bitter, is not well understood. Twelve cheeses were profiled by a trained sensory panel and the five tastes shown to significantly differ in intensity. Sucrose, NaCl, monosodium glutamate, lactic acid, and caffeine were mixed in water and adjusted using a 2^5-1 fractional factorial design (FFD) to reconstruct cheese taste; the optimised construct was compared with a Cheddar cheese to measure similarity for each taste type. The FFD provided knowledge of taste–taste interactions and aided the reconstruction of the taste profile of Cheddar cheese in solution. The reconstructed cheese solution did not significantly differ in overall intensity, saltiness, sourness, umami, and bitterness from the Cheddar cheese based on chi-squared tests. Sweetness was a difficult attribute to adjust due to its relatively low intensity.     

The effect of wild lactic acid bacteria on the production of goat’s milk soft cheese

A blend of four wild strains of lactic acid bacteria, Lactobacillus delbrueckii subsp. bulgaricus, Streptococcus thermophilus, Lactobacillus paracasei subsp. paracasei and Enterococcus faecalis, was used as starter to produce a goat’s milk soft cheese. The cheese was analysed microbiologically, physicochemically and organoleptically through a 30‐day period of ripening. Counts of the starter cultures increased in the first 24 h and then remained stable during ripening. Yeasts and coliforms were found at low numbers initially and gradually decreased to undetectable levels. Micrococcal counts were high throughout ripening. The end product was a soft cheese characterised by a mild, aromatic taste as well as a smooth structure.     

Changes in the microflora of manouri, a traditional Greek whey cheese, during storage

Six batches of manouri cheese were manufactured, and selected microbial groups were counted throughout storage for 20 days at 4°C. The counts of all the microbial groups increased ( P  < 0.05) throughout storage and reached higher levels in cheeses made in summer than in those made in spring. Moreover, the micro-organisms developed better on the cheese surfaces than in the interiors, especially in summer. The pH (6.78–7.33) and salt-in-moisture content (2.53–3.72) of the cheeses did not seem to affect the growth of the bacteria and yeasts present. The isolates of enterobacteriaceae were mainly Hafnia (68.75%), while the isolates from the Baird–Parker medium were mainly staphylococci. There was a great diversity of yeast species, but Debaryomyces hansenii and Pichia membranefasciens predominated. The isolates of enterobacteriaceae, staphylococci and—to a lesser extent—yeasts were proteolytic, but the free amino acid (N-NH2) content of the cheese did not increase significantly during storage (136.4–225.2 mg/kg glycine equivalent). It also seemed that milk fat was not degraded to any great extent (acid degree value 0.09–0.19) by the lipolytic activity of the strains. The main enzymes detected in selected isolates of enterobacteriaceae were leucine aminopeptidase and phosphohydrolase.     

An assessment of factors characterising the microbiology of Grana Trentino cheese,a Grana‐type cheese

In this study, microbiological aspects of Grana Trentino, a variant of Grana Padano cheese, were defined by plate counts, random amplified polymorphic DNA (RAPD) PCR genotying, 16S rRNA gene sequencing of bacterial isolates and PCR–denaturing gradient gel electrophoresis (PCR–DGGE). Results showed variability in monthly fluctuations of whey culture counts, differences in the diffusion of bacterial genotypes among producers and dairy plant‐specific microbial associations. Moreover, the presence of bacteria not previously reported in this cheese type was highlighted, including coagulase‐negative staphylococci and Lactobacillus sanfranciscensis‐like micro‐organisms.     

Changes in the microbiological and chemical characteristics of an artisanal, low-fat cheese made from raw ovine milk during ripening

Changes in the microbial flora of batzos cheese made from raw ovine milk were studied during ripening. Lactic acid bacteria and Enterobacteriaceae were the predominant groups of micro-organisms. Cheeses manufactured in summer had higher microbial counts than those made in spring, with the exception of staphylococci. Nevertheless, Enterobacteriaceae and coliforms decreased more rapidly in cheese made in summer and counts at the end of storage were lower than those in spring cheese.
Enterococci predominated in the ripened curd of cheese made in spring, whereas lactobacilli were the most abundant lactic acid bacteria in cheese made in summer. Enterococcus faecium was the predominant species in spring, and Lactobacillus paracasei ssp. paracasei predominated in cheese made in summer. The pH of the cheeses was > 5.0 throughout ripening, and NaCl-in-moisture content (> 8.0%) permitted the growth and survival of salt-tolerant micro-organisms. α_s1-Casein degraded at a faster rate than β-casein; both caseins were hydrolysed more rapidly in spring than in summer. The free amino acid content became higher in summer cheese (566.24–3460.25 µg/g of glycine equivalent) than in spring cheese because of the progress of ripening. Moreover, the milk fat of the cheese was degraded more in the summer than in the spring. The results suggest that there could be advantages to using starter cultures and improving the level of hygiene during milk and cheese production in order to eliminate undesirable micro-organisms and standardize cheese quality.     

MICROBIOLOGICAL STUDY OF ARZÚA CHEESE (NW SPAIN) THROUGHOUT CHEESEMAKING AND RIPENING

Changes during production and ripening in the microbial flora of 11 batches of Arzúa, a soft cheese made from raw cow's milk, were investigated. The following microbial groups were counted on the surface and interior of the cheese: total viable count (TVC), lactic acid bacteria (LAB), halotolerant flora, enterococci, proteolytic enterococci, staphylococci, Staphylococcus aureus, Enterobacteriaceae, faecal coliforms, molds, yeasts, Listeria spp. and (in milk) Brucella spp. pH and water activity were also determined. TVC and LAB were, generally, more than 9 log (cfu/g). Enterococci counts increased gradually, reaching values in excess of 6 log units. Halotolerant flora and staphylococci remained practically constant throughout ripening, at 6–8 and 5–7 log units, respectively. Maximum Enterobacteriaceae and faecal coliform counts exceeded 7 and 6 log units, respectively. Brucella spp. were not detected in any of the milk samples. Listeria spp. were detected in four batches, and Listeria monocytogenes in two.     

Using PacBio sequencing to investigate the bacterial microbiota of traditional Buryatian cottage cheese and comparison with Italian and Kazakhstan artisanal cheeses

Traditional fermented dairy foods including cottage cheese have been major components of the Buryatia diet for centuries. Buryatian cheeses have maintained not only their unique taste and flavor but also their rich natural lactic acid bacteria (LAB) content. However, relatively few studies have described their microbial communities or explored their potential to serve as LAB resources. In this study, the bacterial microbiota community of 7 traditional artisan cheeses produced by local Buryatian families was investigated using single-molecule, real-time sequencing. In addition, we compared the bacterial microbiota of the Buryatian cheese samples with data sets of cheeses from Kazakhstan and Italy. Furthermore, we isolated and preserved several LAB samples from Buryatian cheese. A total of 62 LAB strains (belonging to 6 genera and 14 species or subspecies) were isolated from 7 samples of Buryatian cheese. Full-length 16S rRNA sequencing of the microbiota revealed 145 species of 82 bacterial genera, belonging to 7 phyla. The most dominant species was Lactococcus lactis (43.89%). Data sets of cheeses from Italy and Kazakhstan were retrieved from public databases. Principal component analysis and multivariate ANOVA showed marked differences in the structure of the microbiota communities in the cheese data sets from the 3 regions. Linear discriminant analyses of the effect size identified 48 discriminant bacterial clades among the 3 groups, which might have contributed to the observed structural differences. Our results indicate that the bacterial communities of traditional artisan cheeses vary depending on geographic origin. In addition, we isolated novel and valuable LAB resources for the improvement of cottage cheese production.     

Effect of midday pasteurizer washing on thermoduric organisms and their progression through Cheddar cheese manufacturing and ripening

Thermoduric bacteria are known to affect the quality of Cheddar cheese, with manifested defects including slits, weak body, and blowing. Thermoduric bacteria are likely to increase in numbers during cheese-making, as in-process conditions are conducive to proliferation. The present study was conducted to track thermoduric bacterial progression during an 18- to 20-h Cheddar cheese production run and during ripening when the pasteurizer was washed at midway through the production day. This study also correlated a broad range of chemical changes to the growth of thermoduric bacteria during ripening. Three independent cheese trials were performed at 3.5- ± 0.5-mo intervals. Samples were drawn in duplicates at 4 different times of the day: at the start of the run (vat 1), prior to a midday wash of the pasteurizer (vat 20), after the midday wash of the pasteurizer (vat 21), and at the end of the run (vat 42) for raw milk, pasteurized milk, and cheese. Cheeses were also tested during ripening for 6 mo. Results showed that raw milk total bacterial counts comprised 0.24% thermoduric mesophiles (TM) and 0.12% thermoduric thermophiles (TT). The thermoduric thermophilic bacterial counts increased by log₁₀ 1.23 during the pasteurizer run of 9 to 10 h, indicating a buildup of thermoduric thermophilic bacteria during the pasteurization process itself. Midday washing reduced thermophilic counts by log₁₀ 1.36, as evident by pre- and post-midday wash counts. However, a thermophilic buildup during post-midday wash was again noticed near the end of the 20-h run. We found that TT bacteria decreased in the first 60 d of ripening, whereas TM bacteria increased during the same period. However, TT bacteria increased later during 60 to 180 d of ripening. Bacillus licheniformis was the most frequently isolated bacteria in this study and was recovered at all production stages sampled during the cheese-making and ripening. We observed a significant increase in the level of orotic and uric acids in the vat made at the end of the day. No significant difference in the overall chemical composition, proteolysis, sugar, or other organic acids was observed in cheese made at the start versus the end of the production run.