HYGIENIC PARAMETERS, TOXINS AND PATHOGEN OCCURRENCE IN RAW MILK CHEESES

In total, 71 samples of retail raw milk cheeses produced or imported in Belgium and samples of Belgian farmhouse cheeses were examined for cotiforms, β-glucuronidase positive Escherichia coli, Escherichia coli O157 , Staphylococcus aureus, Salmonella spp. , Listeria spp. and Listeria monocytogenes. The presence of staphylococcal enterotoxins was investigated on samples with S. aureus counts higher than 10³ cfu/g. The incidence of coliforms, β-glucuronidase positive E. coli and S. aureus was higher in soft than in blue veined, semi-hard, hard and fresh cheeses. Four mold-ripened soft cheeses were positive for E. coli O157. One of the 4 cheeses was positive for verotoxin VT2. Staphylococcal enterotoxins were detected in 1 soft redsmear cheese, which was positive for L. monocytogenes. L. monocytogenes was also detected in one fresh cheese . Salmonella was not detected in any of the 71 raw milk cheeses.     

Diversity of Staphylococcus species and prevalence of enterotoxin genes isolated from milk of healthy cows and cows with subclinical mastitis

The objectives of this study were to determine the occurrence and diversity of Staphylococcus spp. in milk from healthy cows and cows with subclinical mastitis in Brazil and to examine the profile of enterotoxin genes and some enterotoxins produced by Staphylococcus spp. A total of 280 individual mammary quarter milk samples from 70 healthy cows and 292 samples from 73 cows with subclinical mastitis were collected from 11 farms in the state of São Paulo, Brazil. Staphylococcus spp. were recovered from 63 (22.5%) samples from healthy cows and from 80 samples (27.4%) from cows with mastitis. The presence of Staphylococcus aureus was significantly different between these 2 groups and was more prevalent in the cows with mastitis. The presence of Staphylococcus saprophyticus was also significantly different between these 2 groups, but this organism was more prevalent in healthy cows. No statistically significant differences were observed in the numbers of other staphylococci in milk samples from the 2 groups. The sea gene was the most prevalent enterotoxin gene in both groups. Eight of 15 (53.3%) Staph. aureus carried this gene and all produced the SEA toxin. In the coagulase-negative staphylococci (CNS) group, 61 of 128 (47.5%) had the same gene and just 1 (1.6%) Staphylococcus epidermidis strain produced the enterotoxin in vitro. Because CNS were isolated from both groups of cows and most CNS contained enterotoxin genes but did not produce toxins, the role of CNS in mastitis should be carefully defined.     

Microbial and sensory changes throughout the ripening of Prato cheese made from milk with different levels of somatic cells

The objective of this research was to evaluate the effects of 2 levels of raw milk somatic cell count (SCC) on the composition of Prato cheese and on the microbiological and sensory changes of Prato cheese throughout ripening. Two groups of dairy cows were selected to obtain low-SCC (<200,000 cells/mL) and high-SCC (>700,000 cells/mL) milks, which were used to manufacture 2 vats of cheese. The pasteurized milk was evaluated according to the pH, total solids, fat, total protein, lactose, standard plate count, coliforms at 45°C, and Salmonella spp. The cheese composition was evaluated 2 d after manufacture. Lactic acid bacteria, psychrotrophic bacteria, and yeast and mold counts were carried out after 3, 9, 16, 32, and 51 d of storage. Salmonella spp., Listeria monocytogenes, and coagulase-positive Staphylococcus counts were carried out after 3, 32, and 51 d of storage. A 2 × 5 factorial design with 4 replications was performed. Sensory evaluation of the cheeses from low- and high-SCC milks was carried out for overall acceptance by using a 9-point hedonic scale after 8, 22, 35, 50, and 63 d of storage. The somatic cell levels used did not affect the total protein and salt:moisture contents of the cheeses. The pH and moisture content were higher and the clotting time was longer for cheeses from high-SCC milk. Both cheeses presented the absence of Salmonella spp. and L. monocytogenes, and the coagulase-positive Staphylococcus count was below 1 × 10² cfu/g throughout the storage time. The lactic acid bacteria count decreased significantly during the storage time for the cheeses from both low- and high-SCC milks, but at a faster rate for the cheese from high-SCC milk. Cheeses from high-SCC milk presented lower psychrotrophic bacteria counts and higher yeast and mold counts than cheeses from low-SCC milk. Cheeses from low-SCC milk showed better overall acceptance by the consumers. The lower overall acceptance of the cheeses from high-SCC milk may be associated with texture and flavor defects, probably caused by the higher proteolysis of these cheeses.     

Enumeration of coagulase and thermonuclease-positive Staphylococcus spp. in raw milk and fresh soft cheese: An evaluation of Baird-Parker agar,Rabbit Plasma Fibrinogen agar and the Petrifilm™ Staph Express count system

Staphylococcus spp. are microorganisms that are naturally present in milk and dairy products and are often associated with food-borne diseases outbreaks due to the ability of some strains to produce thermostable enterotoxins. This ability is usually associated with coagulase and thermonuclease production, characteristics that are considered in the microbiological analyses for the control of such microorganisms. The objective of this study was to evaluate the culture media and the methodologies used for the enumeration of coagulase and thermonuclease-positive Staphylococcus spp. in raw milk and fresh soft cheese. Samples of artificially contaminated milk (with coagulase-positive Staphylococcus reference strains) and samples of naturally contaminated raw milk and cheese were submitted for enumeration in Baird-Parker agar (BP), Rabbit Plasma Fibrinogen agar (RPFA) and in the Petrifilm™ Staph Express count system (STX). No significant differences (P > 0.05) were observed between the mean counts obtained in all of the evaluated culture media. RPFA and STX had good correlation indices between the total and typical colony counts as well as with coagulase and the thermonuclease-positive colony counts. Thus, there is a better association between coagulase and thermonuclease production to typical colony morphology developed on these culture media, leading to more accurate and reliable results than with BP, which demonstrated lower correlation indices between these counts.     

Traditional Colonial-type cheese from the south of Brazil: A case to support the new Brazilian laws for artisanal cheese production from raw milk

Artisanal Colonial-type cheese is made from raw milk and is the main cheese produced by rural families of the southern region of Brazil. The aim of this study was to investigate, identify problems, and propose solutions for the current situation of small family farms producing and informally selling artisanal Colonial-type cheese located in the western part of Santa Catarina State in Southern Brazil. A semistructured questionnaire was employed in 12 rural properties to analyze the mode of production. Physical-chemical and microbiological analyses of water, raw milk, and cheese were performed, and it was found that 92, 50, and 100% of the samples, respectively, were outside of the current Brazilian regulatory parameters. None of the cheesemakers involved in this study met the requirements, as established by law, for artisanal cheese production from raw milk. This study concluded that technical support and changes in public policy are needed to ensure the preservation of this artisanal cheese, considering the historical importance and cultural traditions of these local communities and the socioeconomic importance of cheesemaking to family farming. Furthermore, more research on the safety of the cheese produced from raw milk is needed as well as the development of specific microbiological standards for artisanal Brazilian cheeses. Public policies aimed at guaranteeing food safety that formalize the commercialization of these cheeses will increase food security in those communities that currently produce artisanal cheese informally.     

Application of indigenous strains of lactic acid bacteria for semi‐industrial production of autochthonous Montenegrin Njeguši cheese

Most of the traditional dairy products of south‐eastern Europe are produced from unpasteurised milk without addition of selected starter cultures. These artisanal products have an interesting biodiversity of indigenous microbiota. In Montenegro, one of the most popular traditional cheeses is Njegu?i cheese, belonging to the group of hard cheeses. Most of the cheeses are produced from ewes' milk, but cows' milk is also widely used. In Montenegro, there are no existing legal regulations defining the specificities of the production or the raw material used for the production of autochthonous food products. Therefore, the producers make the decision about what type of milk to use for cheese production. One of the priorities of the food producers in Montenegro is to obtain a larger scale, standardised production of traditional food products, which are of good quality and considered safe for the consumers. Therefore, in this study, indigenous strains of lactic acid bacteria isolated from various Montenegrin artisanal dairy products were tested as starter cultures for the production of Njegu?i cheese. Three isolates were selected and used as starter cultures in pilot plant experiments for the production of Njegu?i cheese. When applying an adequate combination of added bacterial strains as starters, the flavour of Njegu?i cheese, produced under pilot plant conditions, was comparable to that of the artisanal origin.     

Microbiota characterization of a Belgian protected designation of origin cheese,Herve cheese,using metagenomic analysis

Herve cheese is a Belgian soft cheese with a washed rind, and is made from raw or pasteurized milk. The specific microbiota of this cheese has never previously been fully explored and the use of raw or pasteurized milk in addition to starters is assumed to affect the microbiota of the rind and the heart. The aim of the study was to analyze the bacterial microbiota of Herve cheese using classical microbiology and a metagenomic approach based on 16S ribosomal DNA pyrosequencing. Using classical microbiology, the total counts of bacteria were comparable for the 11 samples of tested raw and pasteurized milk cheeses, reaching almost 8 log cfu/g. Using the metagenomic approach, 207 different phylotypes were identified. The rind of both the raw and pasteurized milk cheeses was found to be highly diversified. However, 96.3 and 97.9% of the total microbiota of the raw milk and pasteurized cheese rind, respectively, were composed of species present in both types of cheese, such as Corynebacterium casei, Psychrobacter spp., Lactococcus lactis ssp. cremoris, Staphylococcus equorum, Vagococcus salmoninarum, and other species present at levels below 5%. Brevibacterium linens were present at low levels (0.5 and 1.6%, respectively) on the rind of both the raw and the pasteurized milk cheeses, even though this bacterium had been inoculated during the manufacturing process. Interestingly, Psychroflexus casei, also described as giving a red smear to Raclette-type cheese, was identified in small proportions in the composition of the rind of both the raw and pasteurized milk cheeses (0.17 and 0.5%, respectively). In the heart of the cheeses, the common species of bacteria reached more than 99%. The main species identified were Lactococcus lactis ssp. cremoris, Psychrobacter spp., and Staphylococcus equorum ssp. equorum. Interestingly, 93 phylotypes were present only in the raw milk cheeses and 29 only in the pasteurized milk cheeses, showing the high diversity of the microbiota. Corynebacterium casei and Enterococcus faecalis were more prevalent in the raw milk cheeses, whereas Psychrobacter celer was present in the pasteurized milk cheeses. However, this specific microbiota represented a low proportion of the cheese microbiota. This study demonstrated that Herve cheese microbiota is rich and that pasteurized milk cheeses are microbiologically very close to raw milk cheeses, probably due to the similar manufacturing process. The characterization of the microbiota of this particular protected designation of origin cheese was useful in enabling us to gain a better knowledge of the bacteria responsible for the character of this cheese.     

Microbiological characterisation of artisanal farmhouse cheeses manufactured in Scotland

Twenty‐eight Scottish artisanal farmhouse cheeses were examined in respect of 16 microbial groups of significance for food safety and cheese character development. Microbial populations were diverse and although Escherichia coli O157 and Salmonella spp. were not detected the occurrence of potential foodborne pathogens was confirmed in 86% of the samples analysed. Mycobacterium avium subsp. paratuberculosis was detected in 25% of the cheeses tested and some Staphylococcus aureus and the Bacillus cereus isolates were enterotoxigenic. Resistance to methicillin and vancomycin and other clinically important antibiotics was detected in some S. aureus and Enterococcus strains. The inappropriate labelling of some raw milk cheeses and the consequences of the complexity of the microbial population on isolation media specificity is discussed.     

Characterisation of the technological behaviour of mixtures of mesophilic lactic acid bacteria isolated from traditional cheeses made of raw milk without added starters

In this work, the technological behaviour in milk of a set of Lactococcus lactis strains, alone or in combination with strains of Leuconostoc spp. and Lactobacillus spp. isolated from traditional, raw milk cheeses made without commercial starters, was investigated. Small, mixture‐specific differences during milk fermentation were recorded for growth, milk acidification and production of organic acids, volatile compounds, free amino acids and biogenic amines. Four combinations appropriate for use as dairy starters were tested in pilot‐scale cheese trials. Two mixtures produced cheeses of high flavour and taste quality; these could be confidently used as starter cultures.     

A review of the microbiological hazards of dairy products made from raw milk

This review concentrates on information concerning microbiological hazards possibly present in raw milk dairy products, in particular cheese, butter, cream and buttermilk. The main microbiological hazards of raw milk cheeses (especially soft and fresh cheeses) are linked to Listeria monocytogenes, verocytotoxin-producing Escherichia coli (VTEC), Staphylococcus aureus, Salmonella and Campylobacter. L. monocytogenes, VTEC and S. aureus have been identified as microbiological hazards in raw milk butter and cream albeit to a lesser extent because of a reduced growth potential compared with cheese. In endemic areas, raw milk dairy products may also be contaminated with Brucella spp., Mycobacterium bovis and the tick-borne encephalitis virus (TBEV). Potential risks due to Coxiella burnetii and Mycobacterium avium subsp. paratuberculosis (MAP) are discussed. Pasteurisation ensures inactivation of vegetative pathogenic microorganisms, which increases the safety of products made thereof compared with dairy products made from raw milk. Several control measures from farm to fork are discussed.     

Short communication: Antimicrobial activity of pequi (Caryocar brasiliense) waste extract on goat Minas Frescal cheese presenting sodium reduction

Sodium chloride reduction in foods is a significant focus of the dairy industry; however, it can interfere with dairy product quality. Thus, researchers have carried out studies on alternatives to maintain dairy product safety when presenting reduced NaCl content, such as natural antimicrobial addition. Caryocar brasiliense (pequi) is a fruit with high phenolic compound concentrations in the pulp and peel and known antioxidant and antimicrobial properties. This study aimed to define the optimum stage for pequi waste extract addition during cheese manufacturing in order to maintain and prolong the shelf life of reduced-sodium goat Minas Frescal cheese. Four different goat Minas Frescal cheese treatments were carried out: control cheese (without extract; CC), pequi extract addition to milk (CM), pequi extract addition to mass (CS), and cheese immersion in pequi extract (CIE). The treatments were subjected to microbiological (Staphylococcus spp., Escherichia coli, Enterobacteriaceae, coliforms and fecal coliforms, Lactococcus spp., and lactic acid bacteria counts), textural (hardness and consistency), and instrumental color (luminosity, yellow intensity, red intensity, chroma, hue angle, and total color change) analyses. No Enterobacteriaceae, Staphylococcus spp., E. coli, or coliforms and fecal coliforms were detected during storage for any of the assessed samples, including CC. Regarding texture, all samples presented a trend for decreasing rigidity during storage. In addition, lower luminosity values were also observed in cheeses produced with added pequi extract (CM, CS, and CIE) when compared with CC. All cheeses produced with added pequi were stable regarding all evaluated parameters; however, pequi extract addition to milk (CM) was shown to be more efficient, leading to higher textural parameters and better microbiological quality during storage. Thus, the CM treatment is the most recommended for pequi waste extract addition during Minas Frescal cheese manufacture.     

Microbial safety status of Serro artisanal cheese produced in Brazil

Considering the growing consumption of artisanal foods worldwide, we aimed to evaluate the microbial safety of Serro artisanal cheese (SAC), produced in Minas Gerais State, Brazil. This cheese is produced with raw milk using 1 of 2 natural starter cultures: “pingo” and “rala.” A total of 53 SAC samples (pingo = 8; rala = 45) were obtained from different farmers and subjected to conventional and molecular assays to detect and enumerate Listeria monocytogenes, Salmonella spp., coagulase-positive staphylococci (CPS), diarrheagenic Escherichia coli, Mycobacterium tuberculosis, and Brucella abortus. The SAC samples were also subjected to an ELISA to detect classical staphylococcal enterotoxins (CSE: SEA, SEB, SEC, SED, SEE) and to PCR assays to detect staphylococcal enterotoxin-related genes (sea, seb, sec, sed, see). Coagulase-positive staphylococci isolates were obtained and tested by the same assays to detect their potential in CSE production and presence of CSE-related genes. None of the SAC samples showed any of the screened food-borne pathogens and zoonotic agents, and none showed the presence of CSE by phenotypic and genotypic approaches. Despite the absence of microbial hazards, mean counts of CPS in SAC samples were 5.2 log cfu/g (pingo starter) and 4.6 log cfu/g (rala starter), indicating poor hygiene practices during production. None of the tested CPS isolates (n = 116) produced CSE or presented CSE-related genes. Despite the relative microbial safety, hygienic conditions during SAC production must be improved to meet official guidelines established in Brazil.     

How three adventitious lactic acid bacteria affect proteolysis and organic acid production in model Portuguese cheeses manufactured from several milk sources and two alternative coagulants

Model cheeses were manufactured according to a full factorial experimental design to help shed light on the individual and combined roles played by 3 native lactic acid bacteria (Lactococcus lactis ssp. lactis, Lactobacillus brevis, and Lactobacillus plantarum) upon proteolysis and organic acid evolution in cheese. The model cheeses were manufactured according to a generally representative Portuguese artisanal protocol, but the (ubiquitous) adventitious microflora in the cheesemaking milk were removed via sterilization before manufacture; therefore, the specific effects of only those lactic acid bacteria selected were monitored. In addition, 2 types of coagulant (animal and plant) and 3 types of cheesemaking milk (cow, sheep, and goat) were assessed to determine their influence on the final characteristics of the model cheeses. The nature of the coagulant appeared to be essential during the first stage of proteolysis as expected, whereas the contribution of those bacteria to the pools of total free AA and organic acids was crucial afterward. This was especially so in terms of the differences observed in the metabolisms of lactic acid (in the case of Lactococcus spp.) as well as acetic and citric acids (in the case of Lactobacillus spp.).     

A BELGIAN SURVEY OF HYGIENE INDICATOR BACTERIA AND PATHOGENIC BACTERIA IN RAW MILK AND DIRECT MARKETING OF RAW MILK FARM PRODUCTS

To monitor the bacteriological quality of raw milk and raw milk farm products, 143 samples of raw farm milk and 100 samples of raw milk farm products, 64 butters, 9 yogurts, 16 cheeses, 7 ice creams and 4 fresh cheeses, produced in Belgium were examined for coliforms, β-glucuronidase positive Escherichia coli, verotoxin producing Escherichia coli O157 , Staphylococcus aureus, Salmonella spp. , Listeria spp. and Listeria monocytogenes. The results were compared with the threshold and maximum values of the EC directive 92/46/EC or the maximum values of the Belgian Order of Council from September 3, 2000. The presence of staphylococcal enterotoxins was investigated on samples with S. aureus counts higher than the legal threshold values mentioned in the EC directive or, if not regulated in the directive, higher than the maximum value mentioned in the Belgian Order of Council. The obtained results for the hygiene-indicators coliforms, β-glucuronidase positive E. coli and S. aureus in the raw milk samples were comparable with most other industrialized countries. Compared to a prevalence of 0.7% and 6.3% for, respectively , E. coli O157 and L. monocytogenes, no Salmonella was found in the 25 g raw milk farm samples. The isolated E. coli O157 strain was confirmed to be verotoxigenic; it was positive for VT2 , eaeA and hly A. In butter not only a prevalence of 18.7% for L. monocytogenes in 25 g was found but also the maximum values for the hygiene-indicators mentioned in the Belgian Order of Council were often exceeded. No significant difference was found between the count of hygiene-indicators and the presence of Listeria spp. as well in raw milk as in raw milk butter. The bacteriological quality of on-farm made raw milk butter suggest that suitable hygienic conditions are not always provided. One of the 7 ice cream samples contained L. monocytogenes in 25 g.     

Acute-phase inter-alpha-trypsin inhibitor heavy chain 4 (ITIH4) levels in serum and milk of cows with subclinical mastitis caused by Streptococcus species and coagulase-negative Staphylococcus species

The aim of the study was to investigate the concentrations of acute-phase inter-α-trypsin inhibitor heavy chain 4 (ITIH4) in serum and milk of cows with subclinical mastitis caused by Streptococcus spp. (STR) and coagulase-negative Staphylococcus spp. (CNS) and healthy cows. The blood and milk samples were obtained from 60 mid-lactation, multiparous Holstein-Friesian cows from 7 herds in the Lublin region of Poland. In the milk samples from 40 cows with subclinical mastitis, Streptococcus spp. and CNS were isolated. The ITIH4 was significantly higher in serum of cows with subclinical mastitis caused both by STR and CNS compared with healthy cows. One hundred percent of animals infected with Streptococcus spp. and 89% of animals infected with Staphylococcus spp. showed ITIH4 concentration in sera higher than 0.5 mg/mL. The concentration of ITIH4 in milk also was significantly higher in cows with subclinical mastitis caused by Streptococcus spp. and Staphylococcus spp. compared with the control group. Seventy percent of cows infected by STR and CNS showed ITIH4 concentration in milk higher than 2.5 μg/mL. Milk ITIH4 concentration higher than 5 µg/mL was found in 55% of animals infected with Streptococcus spp. and in 40% of animals infected with Staphylococcus spp. No statistically significant differences were observed in ITIH4 concentrations both in serum and in milk between the studied unhealthy animal groups. These results suggest that ITIH4 may be used in the future as a novel diagnostic marker in serum and in milk of subclinical mastitis in cows.     

Short communication: High frequency of β-lactam-resistant Staphylococcus aureus in artisanal coalho cheese made from goat milk produced in northeastern Brazil

Reports of β-lactam-resistant Staphylococcus aureus in artisanal goat cheese are increasing, and this phenomenon is relevant to public health. The objective of the present study was to determine the prevalence of S. aureus strains carrying the blaZ and mecA resistance genes, as well as the genes encoding the staphylococcal enterotoxins SEA, SEB, SEC, SED, SEE, and TSST-1 in artisanal coalho cheese made from goat milk produced in northeastern Brazil. We used biochemical and molecular tests to characterize 54 S. aureus isolates found in artisanal coalho cheese collected from commercial establishments producing animal products in 11 municipalities of Pernambuco State, Brazil. A PCR analysis revealed that 42.6% (23/54) of the isolates were positive for the blaZ gene, and 7.4% (4/54) were resistant to methicillin by phenotypic testing. We did not detect mecA or any genes encoding enterotoxins. The presence of S. aureus carriers of the blaZ gene and the identification of methicillin-resistant S. aureus strains are of concern for the health of consumers of this type of cheese.     

Utilization of microfiltration or lactoperoxidase system or both for manufacture of Cheddar cheese from raw milk

The objective of the present study was to determine if application of microfiltration (MF) or raw milk lactoperoxidase system (LP) could reduce the risk of foodborne illness from Escherichia coli in raw milk cheeses, without adversely affecting the overall sensory acceptability of the cheeses. Escherichia coli K12 was added to raw milk to study its survival as a non-pathogenic surrogate organism for pathogenic E. coli. Five replications of 6 treatments of Cheddar cheese were manufactured. The 6 treatments included cheeses made from pasteurized milk (PM), raw milk (RM), raw milk inoculated with E. coli K12 (RME), raw milk inoculated with E. coli K12 + LP activation (RMELP), raw milk inoculated with E. coli K12 + MF (MFE), and raw milk inoculated with E. coli K12 + MF + LP activation (MFELP). The population of E. coli K12 was enumerated in the cheese milks, in whey/curds during cheese manufacture, and in final Cheddar cheeses during ripening. Application of LP, MF, and a combination of MF and LP led to an average percentage reduction of E. coli K12 counts in cheese milk by 72, 88, and 96%, respectively. However, E. coli K12 populations significantly increased during the manufacture of Cheddar cheese for the reasons not related to contamination. The number of E. coli K12, however, decreased by 1.5 to 2 log cycles during 120 d of ripening, irrespective of the treatments. The results suggest that MF with or without LP significantly lowers E. coli count in raw milk. Hence, if reactivation of E. coli during cheese making could be prevented, MF with or without LP would be an effective technique for reducing the counts of E. coli in raw milk cheeses. The cheeses were also analyzed for proteolysis, starter and nonstarter lactic acid bacteria (NSLAB), and sensory characteristics during ripening. The concentration of pH 4.6 soluble nitrogen at 120 d was greater in PM cheese compared with the other treatments. The level of 12% trichloroacetic acid-soluble nitrogen at 120 d was greater in RM, RME, and RMELP cheeses compared with PM, MFE, and MFELP cheeses. This could be related to the fact that cheeses made from raw milk with or without LP (RM, RME, and RMELP) had greater levels of NSLAB compared with PM, MFE, and MFELP cheeses. Cheeses at 60 d, as evaluated by 8 trained panelists, did not differ in bitterness, pastiness, or curdiness attributes. Cheeses at 120 d showed no differences in acid-taste, bitterness, or curdiness attributes. Sensory analysis at 60 d showed that PM and MFELP cheeses had greater overall sensory acceptability than RM and RME cheeses. The overall sensory acceptability of the cheeses at 120 d showed that PM, MFE, and MFELP cheeses were more acceptable than RM and RME cheeses.     

Characterization of the lactic acid bacteria in ewe's milk and cheese from northwest Argentina

Indigenous lactic acid bacteria in ewe's milk and artisanal cheese were studied in four samples of fresh raw milk and four 1-month-old cheeses from the provinces of northwest Argentina. Mean growth counts on M17, MRS, and MSE agar media did not show significant differences (P < 0.05) in raw milk and cheeses. Isolates of lactic acid bacteria from milk were identified as Enterococcus (48%), lactococci (14%), leuconostocs (8%), and lactobacilli (30%). All lactococci were identified as Lactococcus lactis (subsp. lactis and subsp. cremoris). Lactobacilli were identified as Lactobacillus plantarum (92%) and Lactobacillus acidophilus (8%). Enterococci (59%) and lactobacilli (41%) were isolated from cheeses. L. plantarum (93%), L. acidophilus (5%), and Lactobacillus casei (2%) were most frequently isolated. L. lactis subsp. lactis biovar diacetylactis strains were considered as fast acid producers. L. lactis subsp. cremoris strains were slow acid producers. L. plantarum and L. casei strains identified from the cheeses showed slow acid production. The majority of the lactobacilli and Lactococcus lactis strains utilized citrate and produced diacetyl and acetoin in milk. Enzyme activities (API-ZYM tests) of lactococci were low, but activities of L. plantarum strains were considerably higher. The predominance of L. plantarum in artisanal cheese is probably important in the ripening of these cheeses due to their physiological and biochemical characteristics.     

Foci of contamination of Listeria monocytogenes in different cheese processing plants

Listeria monocytogenes is a ubiquitous bacterium widely distributed in the environment that can cause a severe disease in humans when contaminated foods are ingested. Cheese has been implicated in sporadic cases and in outbreaks of listeriosis worldwide. Environmental contamination, in several occasions by persistent strains, has been considered an important source of finished product contamination. The objectives of this research were to (i) evaluate the presence of L. monocytogenes within the factory environments and cheeses of three processing plants, artisanal producer of raw ewe's milk cheeses (APC), small-scale industrial cheese producer (SSI) and industrial cheese producer (ICP) each producing a distinct style of cheese, all with history of contamination by L. monocytogenes (ii) and identify possible sources of contamination using different typing methods (arsenic and cadmium susceptibility, geno-serotyping, PFGE). The presence of markers specific for 3 epidemic clones (ECI–ECIII) of L. monocytogenes was also investigated. Samples were collected from raw milk (n = 179), whey (n = 3), cheese brining solution (n = 7), cheese brine sludge (n = 505), finished product (n = 3016), and environment (n = 2560) during, at least, a four-year period. Listeria monocytogenes was detected in environmental, raw milk and cheese samples, respectively, at 15.4%, 1.1% and 13.6% in APC; at 8.9%, 2.9% and 3.4% in SSI; and at 0%, 21.1% and 0.2% in ICP. Typing of isolates revealed that raw ewe's milk and the dairy plant environment are important sources of contamination, and that some strains persisted for at least four years in the environment. Although cheeses produced in the three plants investigated were never associated with any case or outbreak of listeriosis, some L. monocytogenes belonging to specific PFGE types that caused disease (including putative epidemic clone strains isolated from final products) were found in this study.     

Factors affecting consumers' preferences for and purchasing decisions regarding pasteurized and raw milk specialty cheeses

Eight hundred ninety consumers at a local food festival were surveyed about their specialty cheese purchasing behavior and asked to taste and rate, through nonforced choice preference, 1 of 4 cheese pairs (Cheddar and Gouda) made from pasteurized and raw milks. The purpose of the survey was to examine consumers’ responses to information on the safety of raw milk cheeses. The associated consumer test provided information about specialty cheese consumers’ preferences and purchasing behavior. Half of the consumers tested were provided with cheese pairs that were identified as being made from unpasteurized and pasteurized milk. The other half evaluated samples that were identified only with random 3-digit codes. Overall, more consumers preferred the raw milk cheeses than the pasteurized milk cheeses. A larger portion of consumers indicated preferences for the raw milk cheese when the cheeses were labeled and thus they knew which samples were made from raw milk. Most of the consumers tested considered the raw milk cheeses to be less safe or did not know if raw milk cheeses were less safe. After being informed that the raw milk cheeses were produced by a process approved by the FDA (i.e., 60-d ripening), most consumers with concerns stated that they believed raw milk cheeses to be safe. When marketing cheese made from raw milk, producers should inform consumers that raw milk cheese is produced by an FDA-approved process.