Microbiological quality of retail cheeses made from raw, thermized or pasteurized milk in the UK

Two studies of retail fresh, ripened and semi-hard cheeses made from raw, thermized or pasteurized milk were undertaken in the UK during 2004 and 2005 to determine the microbiological quality of these products. Using microbiological criteria in European Commission Recommendations 2004/24/EC and 2005/175/EC, 2% of both raw, thermized (37/1819 samples) and pasteurized (51/2618 samples) milk cheeses were of unsatisfactory quality. Raw or thermized milk cheeses were of unsatisfactory quality due to levels of Staphylococcus aureus at 10(4)cfu g(-1), Escherichia coli at 10(5)cfu g(-1), and/or Listeria monocytogenes at 10(2)cfu g(-1), whereas pasteurized milk cheeses were of unsatisfactory quality due to S. aureus at 10(3)cfu g(-1) and/or E. coli at 10(3)cfu g(-1). Salmonella was not detected in any samples. Cheeses were of unsatisfactory quality more frequently when sampled from premises rated as having little or no confidence in management and control systems, and stored/displayed at above 8 degrees C. Raw or thermized milk cheeses were also more likely to be of unsatisfactory quality when they were unripened types, and pasteurized milk cheeses when they were: semi-hard types; from specialist cheese shops or delicatessens; cut to order. These results emphasize the need for applying and maintaining good hygiene practices throughout the food chain to prevent contamination and/or bacterial growth. Labelling of cheeses with clear information on whether the cheese was prepared from raw milk also requires improvement.     

Occurrence of foodborne pathogens in Irish farmhouse cheese

Food safety is a critical factor in the production of farmhouse cheese. In Ireland the varieties of farmhouse cheese produced reflect a much broader range than those produced commercially and some of these cheese varieties are associated with greater microbiological risk. These include cheese produced from unpasteurised milk and soft ripened cheese such as mould or smear-ripened cheeses which have high pH and relatively short ripening times. The aim of this study was to determine the microbiological quality of farmhouse cheeses in Ireland. Three hundred and fifty one cheese samples, from 15 cheese producers, were analysed for microbiological quality on a monthly basis throughout the year. The analyses included enumeration of Escherichia coli, Staphylococcus aureus and Listeria monocytogenes (using the relevant agars) and enrichment for L. monocytogenes. The cheeses selected were produced from ovine, caprine and bovine milk. Both unpasteurised and pasteurised milk cheeses were sampled and these included hard, semi-hard and soft cheeses, internal/external mould-ripened and smear-ripened cheeses and the cheeses represented different geographic regions. Of the cheeses tested, 94% were free of L. monocytogenes, all were within the EU limits for E. coli and only one cheese variety had S. aureus levels above the recommended numbers for the first 6 months of the year. Due to a modified production process the numbers were within the guidelines for the second six months. The results indicate that Irish farmhouse cheeses are of a high microbiological quality.     

Influence of Australian Native Herbs on the Maturation of Vacuum-packed Cheese

The composition, microbiology and biochemistry of semi-hard cheeses flavoured with native mint, lemon myrtle and bush tomato (BT) were compared with unflavoured (control) cheese during a 90-day period of maturation. Moisture, protein and salt levels of all cheeses were similar and did not change during maturation. However, the fat content of control cheese was significantly higher than that of the flavoured cheeses while the pH of cheese flavoured with BT was consistently lower throughout maturation. Total viable organisms, Lactobacillus and Lactococcus counts were between 10⁶ and 10⁷ colony forming units (cfu)/g cheese for all cheeses. Yeast and mould count was <10² cfu/g cheese throughout the maturation of all cheeses except in the cheese flavoured with BT which was >10³ cfu/g cheese. Biochemical indices of proteolysis and lipolysis increased with the extent of maturation in all cheeses but were most pronounced in the BT-flavoured cheese. The capillary electrophoretic profile of this cheese also indicated a more extensive hydrolysis of both α s- and β -caseins. The microbiological quality of BT appeared to have exerted a very significant influence on cheese properties.     

The effects of starter culture on chemical composition, microbiological and sensory characteristics of Turkish Kaşar Cheese during ripening

Kaşar cheese samples were produced from raw milk and starter culture-added pasteurized milk. Chemical, microbiological and organoleptic properties of kaşar cheeses were analysed at certain times during the ripening periods (on the 1st, 7th, 15th, 30th, 60th, 90th days). Generally, chemical parameters were not affected by starter culture. The pH, ripening index, water-soluble nitrogen and non-protein nitrogen did not show significant differences between the cheese samples. The addition of starter affected the microbiological quality of the cheeses. Starter culture-added kaşar cheeses contained low levels of total aerobic mesophilic bacteria, moulds and yeasts, and coliforms, and achieved higher organoleptic scores than those of cheeses made from raw milk. The starter cultures contributed to acidity and microbial quality of the cheese.     

Behaviour of Listeria monocytogenes during the manufacture and ripening of Manchego and Chihuahua Mexican cheeses

The ability of Listeria monocytogenes to survive the Mexican Manchego and Chihuahua cheese-making processes and its persistence during the ripening stages of both cheeses was examined. Commercial pasteurized and homogenized whole milk was inoculated with Listeria monocytogenes (strain ATCC 19114) to a level between 2 x 10(6) and 9 x 10(6) CFU/ml. The milk was used to make Mexican Manchego and Chihuahua cheeses in a 25-l vat. Mexican Manchego cheese was ripened for 5 days and Chihuahua cheese for 6 weeks at 12 degrees C and 85% RH. Listeria present in the cheese was enumerated by diluting samples in sterile 0.1% peptone water and plating on Oxford agar. Duplicate samples were taken at each step of the manufacturing process. During the first week of ripening samples were taken daily from both cheeses. For Chihuahua cheese, samples were taken weekly after the first week of the ripening stage. During the manufacture of Mexican Manchego cheese, Listeria counts remained relatively constant at 10(6) CFU/ml, while with Chihuahua cheese there was a one log decrease in numbers (10(6) to 10(5) CFU/ml). After pressing both curds overnight, numbers of bacteria decreased in Mexican Manchego cheese to 8.2 x 10(5) but increased in Chihuahua cheese from 1.7 x 10(5) to 1.2 x 10(6) CFU/ml. During the ripening stage, counts of Listeria remained constant in both cheeses. However, since the Chihuahua cheese ripening stage is about 6 weeks, the number of bacteria decreased from 2 x 10(6) to 4 x 10(4) CFU/g. The results show that Listeria monocytogenes is able to survive the manufacture and ripening processes of both Mexican cheeses.     

Short communication: Assessing antihypertensive activity in native and model Queso Fresco cheeses

Hispanic-style cheeses are one of the fastest growing varieties in the United States, making up approximately 2% of the total cheese production in this country. Queso Fresco is one of most popular Hispanic-style cheeses. Protein extracts from several varieties of Mexican Queso Fresco and model Queso Fresco were analyzed for potential antihypertensive activity. Many Quesos Frescos obtained from Mexico are made from raw milk and therefore the native microflora is included in the cheese-making process. Model Queso Fresco samples were made from pasteurized milk and did not utilize starter cultures. Water-soluble protein extracts from 6 Mexican Quesos Frescos and 12 model cheeses were obtained and assayed for their ability to inhibit angiotensin-converting enzyme, implying potential as foods that can help to lower blood pressure. All model cheeses displayed antihypertensive activity, but mainly after 8 wk of aging when they were no longer consumable, whereas the Mexican samples did display some angiotensin-converting enzyme inhibitory action after minimal aging.     

Heavy metals in cow’s milk and cheese produced in areas irrigated with waste water in Puebla,Mexico

The aim of this work was to determine Ni, Cr, Cu, Zn, Pb, and As levels in raw milk and Oaxaca and ranchero type cheeses, produced in areas irrigated with waste water from Puebla in Mexico. Milk results showed a mean Pb level of 0.03 mg kg^?1, which is above the maximum limit as set by Codex Alimentarius and the European Commission standards. For As a mean value of 0.12 mg kg^?1 in milk was obtained. Mean As and Pb levels in milk were below the Mexican standard. Milk whey and ranchero cheese had mean Pb levels of 0.07 and 0.11 mg kg^?1, respectively. As was higher in Oaxaca and ranchero cheese at 0.17 and 0.16 mg kg^?1, respectively. It was concluded that cheeses made from cow’s milk from areas irrigated with waste water are contaminated with Pb and As, which may represent a health risk.     

Microbiological changes throughout ripening of goat cheese made from raw,pasteurized and high-pressure-treated milk

The bacteriological quality during ripening of raw (RA), pasteurized (PA; 72°C, 15 s) and pressure-treated (PR; 500 MPa, 20°C, 15 min) goat milk assessed by enumeration of total bacteria, psychrotrophic bacteria, Enterobacteriaceae, lactobacilli, enterococci, Micrococcaceae and lactococci was evaluated. The high pressure treatment applied was as efficient as pasteurization in reducing the bacterial population of milk. Experimental cheeses were made from RA, PA and PR milks to study the microbial population during ripening. Lactobacilli and lactococci were the predominant microbiota present during ripening in all the cheeses. There were no differences in numbers of starter bacteria during ripening. However, lactobacilli counts for RA milk cheese were significantly higher than for PA and PR cheeses in all the ripening stages studied. Micrococcaceae and enterococci remained at a secondary level, and no differences were observed between cheeses at the end of ripening. On the other hand, the number of Enterobacteriaceae decreased during ripening, but faster in PR milk cheese than in PA and RA milk cheeses. The results of this study suggest that goat cheese made from PR milk had similar microbiological characteristics to PA milk cheeses.     

Some physicochemical, microbiological, and sensory properties of tulum cheese produced from ewe's milk via a modified method

The purpose of this research was to investigate an alternative way to manufacture Erzincan tulum cheese in order to shorten production time and improve food safety. By adding 0.5% starter culture ( Streptococcus salivarius ssp. thermophilus and Lactobacillus delbrueckii ssp. bulgaricus cultures at a 1 : 1 ratio) to pasteurized ewe's milk (65°C for 30 min), the required time for manufacturing Erzincan tulum cheese was shortened from the traditional 10–12 days to 2 days. The cheeses manufactured with the modified method were ripened in three different packaging materials: goatskin, plastic, and ceramic. Physicochemical, microbiological, and sensory properties of the Erzincan tulum cheese were obtained during the ripening period at intervals of 2, 30, 60, and 90 days, and compared with those properties of samples manufactured by the traditional method. Significant microbiological and physicochemical differences were found between the modified samples and the traditional samples ( P <  0.01). However, the modified samples and the traditional samples were statistically similar in sensory properties to the exception of the modified sample packaged in plastic.     

Fate of Staphylococcus aureus in cheese treated by ultrahigh pressure homogenization and high hydrostatic pressure

We evaluated the influence of ultrahigh pressure homogenization (UHPH) treatment applied to milk containing Staphylococcus aureus CECT 976 before cheese making, and the benefit of applying a further high hydrostatic pressure (HHP) treatment to cheese. The evolution of Staph. aureus counts during 30 d of storage at 8°C and the formation of staphylococcal enterotoxins were also assessed. Milk containing approximately 7.3 log₁₀ cfu/mL of Staph. aureus was pressurized using a 2-valve UHPH machine, applying 330 and 30 MPa at the primary and the secondary homogenizing valves, respectively. Milk inlet temperatures (T_in) of 6 and 20°C were assayed. Milk was used to elaborate soft-curd cheeses (UHPH cheese), some of which were additionally submitted to 10-min HHP treatments of 400 MPa at 20°C (UHPH+HHP cheese). Counts of Staph. aureus were measured on d 1 (24 h after manufacture or immediately after HHP treatment) and after 2, 15, and 30 d of ripening at 8°C. Counts of control cheeses not pressure-treated were approximately 8.5 log₁₀ cfu/g showing no significant decreases during storage. In cheeses made from UHPH treated milk at T_in of 6°C, counts of Staph. aureus were 5.0 ± 0.3 log₁₀ cfu/g at d 1; they decreased significantly to 2.8 ± 0.2 log₁₀ cfu/g on d 15, and were below the detection limit (1 log₁₀ cfu/g) after 30 d of storage. The use of an additional HHP treatment had a synergistic effect, increasing reductions up to 7.0 ± 0.3 log₁₀ cfu/g from d 1. However, for both UHPH and UHPH+HHP cheeses in the 6°C T_in samples, viable Staph. aureus cells were still recovered. For samples of the 20°C T_in group, complete inactivation of Staph. aureus was reached after 15 d of storage for both UHPH and UHPH+HHP cheese. Staphylococcal enterotoxins were found in controls but not in UHPH or UHPH+HHP treated samples. This study shows a new approach for significantly improving cheese safety by means of using UHPH or its combination with HHP.     

Kluyveromyces lactis but not Pichia fermentans used as adjunct culture modifies the olfactory profiles of Cantalet cheese

Yeasts are commonly detected in cheese. Two yeast species, Kluyveromyces lactis and Pichia fermentans, were isolated at high populations from raw-milk Cantal cheese, a French Protected Denomination of Origin hard cheese. To investigate the interest of these 2 species as adjunct cultures to promote flavor development of Cantalet cheese, they were added at 10⁵ cfu/mL to microfiltered milk. The global microbiological, biochemical, and flavor changes induced by the presence of the yeasts in cheese were determined. Adjunct yeasts were present at 10⁶ cfu/g in curd, declined to 10⁴ to 10⁵ cfu/g in cheese, and did not influence gross composition, content of free amino acids, or content of free fatty acids. By using 8-way gas chromatography-olfactometry in parallel with gas chromatography-mass spectrometry, 30 odorous compounds of Cantalet cheese were identified. The olfactory profiles of K. lactis cheeses contained significantly greater levels of 8 odorous compounds (ethanol, ethyl hexanoate, 4 aldehydes, and 2 branched-chain acids) compared with the control and P. fermentans cheeses. Sensory analysis of cheeses flavor discriminated K. lactis cheeses on only 2 attributes (acetaldehyde and alcohol odors). This study shows that yeast contribution is species-specific and that K. lactis, at a population of 10⁶ viable cells/g, can influence Cantalet cheese flavor.     

Effect of different salting technologies on the chemical and microbiological characteristics of PDO Pecorino Siciliano cheese

The present work was carried out to evaluate the effect of two salting technologies [dry salting (DS) and the combined dry-brine salting (DBS)] on the chemico-physical and microbiological characteristics of PDO Pecorino Siciliano cheeses of different final weight (6 and 12 kg). Dry matter was significantly influenced by both salting process and final size. Twelve kilogram cheeses treated by DBS showed higher protein content with higher soluble nitrogen per cent than 6 kg cheeses. Salt content was in the range 3.1–4.0% on dry matter. The colour did not show significant differences for any of the factors, but 12 kg cheeses subjected to DS showed higher yellow index than the other cheeses. The resistance at 30% of strain was influenced by cheese size, with 6 kg cheeses showing higher resistances than 12 kg cheeses. All cheeses were dominated by coccus LAB, but pseudomonads and Enterobacteriaceae showed comparable levels of about 10⁵ cfu/g. Significant microbiological differences were evidenced only for enterococci and yeasts concerning the final cheese size. Thirteen species of LAB, belonging to five genera (Enterococcus, Lactobacillus, Lactococcus, Pediococcus and Streptococcus), were identified, but several spoilage/pathogenic species were also identified, especially Pseudomonas putida, Citrobacter freundii and Stenotrophomonas maltophilia. LAB isolates were preliminary evaluated for their physiological characteristics in view of developing autochthonous starters to improve the microbiological quality of PDO Pecorino Siciliano cheese.     

Influence of the starter culture on the microbiological and sensory characteristics of ewe's cheese

Changes which take place in the sensory characteristics of cheeses during ripening are influenced by different factors, involving rennet, starter culture and adventitious contamination of the cheese by non-starter lactic acid bacteria. The objective of this work was to study the influence of the starter on sensory and microbiological ewe's cheese properties during ripening time. Four batches (two with starter added and two without) were manufactured. Milk and cheeses at different stages of ripening were analysed. Cheeses manufactured without adding starter showed a significantly higher level of mesophilic aerobic microflora, lactobacilli, facultatively heterofermentative lactobacilli and enterococci (indigenous microflora) than cheeses manufactured with starter. This study has also shown that adding or not adding starter affects the flavour profile of the cheese. Cheeses with starter added showed greater intensity of the following attributes: refreshing, astringent, sweet; and received lower scores on bitterness. With respect to texture, the said cheeses develop a more homogenous texture and greater elasticity throughout ripening.     

Short communication: characterization of microflora in Mexican Chihuahua cheese

This work was performed to identify the bacterial species present in 10 Chihuahua cheeses obtained from commercial producers in Mexico using 16S rRNA gene analysis. As expected, some of the agar media initially used for isolation were not very selective, supporting the growth of several unrelated bacterial species. Sequence analysis identified potential pathogens, including Escherichia coli and Staphylococcus aureus, in all raw milk samples and 2 pasteurized milk samples. Streptococcus thermophilus and Lactococcus lactis ssp. lactis were identified in 9 and 6 samples, respectively, and would serve as acidifying agents during cheese production. Lactobacilli were identified in all cheeses, with the most prevalent being Lactobacillus plantarum identified in 7 raw milk and 1 pasteurized milk cheeses. Leuconostoc mesenteroides and Streptococcus macedonicus were identified in 4 raw milk cheeses and both were present in all pasteurized milk samples, suggesting that they may play a role in the development of traditional Chihuahua cheese attributes.     

Nonstarter Lactobacillus strains as adjunct cultures for cheese making: in vitro characterization and performance in two model cheeses

Nonstarter lactic acid bacteria are the main uncontrolled factor in today's industrial cheese making and may be the cause of quality inconsistencies and defects in cheeses. In this context, adjunct cultures of selected lactobacilli from nonstarter lactic acid bacteria origin appear as the best alternative to indirectly control cheese biota. The objective of the present work was to study the technological properties of Lactobacillus strains isolated from cheese by in vitro and in situ assays. Milk acidification kinetics and proteolytic and acidifying activities were assessed, and peptide mapping of trichloroacetic acid 8% soluble fraction of milk cultures was performed by liquid chromatography. In addition, the tolerance to salts (NaCl and KCl) and the phage-resistance were investigated. Four strains were selected for testing as adjunct cultures in cheese making experiments at pilot plant scale. In in vitro assays, most strains acidified milk slowly and showed weak to moderate proteolytic activity. Fast strains decreased milk pH to 4.5 in 8 h, and continued acidification to 3.5 in 12 h or more. This group consisted mostly of Lactobacillus plantarum and Lactobacillus rhamnosus strains. Approximately one-third of the slow strains, which comprised mainly Lactobacillus casei, Lactobacillus fermentum, and Lactobacillus curvatus, were capable to grow when milk was supplemented with glucose and casein hydrolysate. Peptide maps were similar to those of lactic acid bacteria considered to have a moderate proteolytic activity. Most strains showed salt tolerance and resistance to specific phages. The Lactobacillus strains selected as adjunct cultures for cheese making experiments reached 10⁸ cfu/g in soft cheeses at 7 d of ripening, whereas they reached 10⁹ cfu/g in semihard cheeses after 15 d of ripening. In both cheese varieties, the adjunct culture population remained at high counts during all ripening, in some cases overcoming or equaling primary starter. Overall, proximate composition of cheeses with and without added lactobacilli did not differ; however, some of the tested strains continued acidifying during ripening, which was mainly noticed in soft cheeses and affected overall quality of the products. The lactobacilli strains with low acidifying activity showed appropriate technological characteristics for their use as adjunct cultures in soft and semihard cheeses.     

Non-lactic acid, contaminating microbial flora in ready-to-eat foods: a potential food-quality index

The bacteriological profile of 87 samples of commercially available ready-to-eat (RTE) dairy and meat-products, packaged sandwiches and salads was obtained by testing for aerobic colony count, for lactic acid bacterial (LAB) count, for the presence and the extent of non-LAB microflora (contaminating microflora), and by testing for certain food-borne pathogens. The pathogens Listeria monocytogenes, Salmonella spp. and sulfite-reducing clostridia were not detected in any of the analysed samples. Whereas only three samples (3.4%) were deemed unacceptable for consumption for exceeding the established pathogen tolerance levels (for Staphylococcus aureus and Escherichia coli), several samples were found to contain non-lactic acid contaminating microflora of considerable magnitude. The log10 cfu g(-1) counts for contaminating microflora in the food categories examined were as follows: hard cheeses 4.85 (SD 1.17); semi-hard cheeses 5.39 (SD 1.37); soft cheeses 5.13 (SD 1.03); whey cheeses 6.55 (1.24); fermented meat-products 4.18 (SD 1.48); heat-treated meat-products 3.47 (SD 1.99); salads 3.37 (SD 1.56) and sandwiches 5.04 (SD 0.96). Approximately 1 in every 30 to 80 bacterial cells found on different types of cheeses and salads was a non-LAB microorganism; the respective ratios for fermented meat-products, heat-treated meat-products and sandwiches were 1 in 6, 2.5 and 15. The assessment of the contaminating microflora magnitude at various steps during the manufacture and distribution of RTE foods can serve as an index for monitoring the microbiological quality of the starting materials, the sanitation efficacy during processing and possible temperature abuse during processing, transportation or storage.     

High incidence of Listeria monocytogenes in European red smear cheese

The incidence of Listeria and Listeria monocytogenes in European red smear cheese was determined in order to assess whether the lack of recent outbreaks of listeriosis associated with cheese is due to improved hygenic conditions in the dairies. Out of European red-smear cheese samples of various types, 15.8% contained organisms of the genus Listeria, 6.4% of the samples were contaminated with L. monocytogenes, 10.6% with L. innocua, and 1.2% with L. seeligeri. Six cheese samples contained two or more Listeria species, including at least one L. monocytogenes isolate. The incidences of L. monocytogenes in cheeses from various countries were: Italy 17.4%, Germany 9.2%, Austria 10%, and France 3.3%. Listeria were found most frequently in soft and semi-soft cheese. Eight samples contained more than 100 L. monocytogenes cfu/cm2 cheese surface, 2 samples had counts above 10(4) cfu/cm2 cheese surface. Surprisingly, a higher incidence of L. monocytogenes was observed in cheeses made from pasteurized milk (8.0%) than in cheeses manufactured from raw milk (4.8%). Phage-typing of isolated Listeria strains clearly confirmed that (i) contaminations within dairy plants were persistent over a period of several weeks to months and (ii) that cross-contamination within the dairy plant is and important factor. Comparison of our data with past surveys seems to indicate that contamination of red smear soft cheese with L. monocytogenes has not decreased sufficiently over the past 15 years. It is therefore strongly recommended that these products are monitored carefully by cheese-making companies.     

Microbiological characterization of artisanal Raschera PDO cheese: analysis of its indigenous lactic acid bacteria

The aim of this research was to study the bacterial populations involved in the production of artisanal Raschera PDO cheese (Italian Maritime Alps, northwest Italy) in order to collect preliminary knowledge on indigenous lactic acid bacteria (LAB). A total of 21 samples of Raschera PDO cheese, collected from six dairy farms located in the production area, were submitted to microbiological analysis. LAB were randomly isolated from M17 agar, MRS agar and KAA plates and identified by combining PCR 16S-23S rRNA gene spacer analysis, species-specific primers and 16S rRNA gene sequencing. Biodiversity of Lactococcus lactis subsp. lactis isolates was investigated by RAPD-PCR. LAB microflora showed the highest count values among all microbial groups targeted. They reached counts of 10(9) colony forming unit (cfu)/g in cheese samples after 3 days of salting and 15 days of ripening. Yeast population also showed considerable count values, while enterococci and coagulase-negative cocci (CNC) did not overcome 10(7)cfu/g. L. lactis subsp. lactis was the species most frequently isolated from Raschera PDO samples at all different production stages while in aged cheeses Lactobacillus paracasei was frequently isolated. RAPD-PCR highlighted that isolates of L. lactis subsp. lactis isolated from Raschera PDO were highly homogeneous.     

Effect of Microencapsulation on Viability of Lactobacillus acidophilus LA-5 and Bifidobacterium bifidum BB-12 During Kasar Cheese Ripening

The viability of encapsulated Lactobacillus acidophilus LA-5 and Bifidobacterium bifidum BB-12 by emulsion or extrusion techniques in Kasar cheese was investigated. The microbiological, biochemical and organoleptic properties of cheeses were assessed throughout 90-day storage. Results showed that the viability of probiotic bacteria was maintained to a great extent by microencapsulation. No difference was noted between the two encapsulation techniques with regard to bacterial counts, proteolysis and organoleptical properties of the final products. Scalding caused a drastic decline in the counts of probiotic bacteria in all cheeses. Following scalding, while the numbers of nonencapsulated probiotic bacteria decreased continuously in the control cheese, the numbers of encapsulated bacteria remained well above the threshold for a minimum probiotic effect (10⁷ cfu/g).