Bacteriocinogenic anti-listerial properties and safety assessment of Enterococcus faecium and Lactococcus garvieae strains isolated from Brazilian artisanal cheesemaking environment.

AIMS: This study aimed to prospect and isolate lactic acid bacteria (LAB) from an artisanal cheese production environment, to assess their safety, and to explore their bacteriocinogenic potential against Listeria monocytogenes. METHODS AND RESULTS: Samples were collected from surfaces of an artisanal-cheese production facility and after rep-PCR and 16S rRNA sequencing analysis, selected strains were identified as to be belonging to Lactococcus garvieae (1 strain) and Enterococcus faecium (14 isolates, grouped into three clusters) associated with different environments (worktables, cheese mold, ripening wooden shelves). All of them presented bacteriocinogenic potential against L. monocytogenes ATCC 7644 and were confirmed as safe (γ-hemolytic, not presenting antibiotic resistance, no mucus degradation properties, and no proteolytic or gelatinase enzyme activity). Additionally, cell growth, acidification and bacteriocins production kinetics, bacteriocin stability in relation to different temperatures, pH, and chemicals were evaluated. According to performed PCR analysis all studied strains generated positive evidence for the presence of entA and entP genes (for production of enterocins A and enterocins P, respectively). However, pediocin PA-1 associated gene was recorded only in DNA obtained from E. faecium ST02JL and Lc. garvieae ST04JL. CONCLUSIONS: It is worth considering the application of these safe LAB or their bacteriocins in situ as an alternative means of controlling L. monocytogenes in cheese production environments, either alone or in combination with other antimicrobials.

Replacing animal proteins with plant proteins: Is this a way to improve quality and functional properties of hybrid cheeses and cheese analogs?

The growing emphasis on dietary health has facilitated the development of plant-based foods. Plant proteins have excellent functional attributes and health-enhancing effects and are also environmentally conscientious and animal-friendly protein sources on a global scale. The addition of plant proteins (including soy protein, pea protein, zein, nut protein, and gluten protein) to diverse cheese varieties and cheese analogs holds the promise of manufacturing symbiotic products that not only have reduced fat content but also exhibit improved protein diversity and overall quality. In this review, we summarized the utilization and importance of various plant proteins in the production of hybrid cheeses and cheese analogs. Meanwhile, classification and processing methods related to these cheese products were reviewed. Furthermore, the impact of different plant proteins on the microstructure, textural properties, physicochemical attributes, rheological behavior, functional aspects, microbiological aspects, and sensory characteristics of both hybrid cheeses and cheese analogs were discussed and compared. Our study explores the potential for the development of cheeses made from full/semi-plant protein ingredients with greater sustainability and health benefits. Additionally, it further emphasizes the substantial chances for scholars and developers to investigate the optimal processing methods and applications of plant proteins in cheeses, thereby improving the market penetration of plant protein hybrid cheeses and cheese analogs.

Determination of safety status and probiotic properties of Enterococcus strains isolated from traditional cheeses in Turkey.

AIMS: This study aimed to evaluate the probiotic properties of Enterococcus strains isolated from Turkish traditional cheeses. METHODS AND RESULTS: Fifty-two Enterococcus spp. were taxonomically determined as follows: Enterococcus faecium (26), Enterococcus faecalis (18), Enterococcus durans (6), and Enterococcus italicus (2). The ability of isolates/strains to survive the harsh conditions (acidity and in-vitro gastric solution) of the gastrointestinal tract was established. They also showed auto-aggregation, hydrophobicity, and co-aggregation ability. Hydrophobicities of the strains were found between 0.8%-21%, 0.7%-56%, and 2%-63% for xylene, chloroform, and ethyl acetate, respectively. Autoaggregation values of the Enterococcus strains were 4%-20%, 7%-30%, and 36%-98% after 2, 4, and 24-h incubation, respectively. In this study, the Enterococcus strains tested showed co-aggregation ability with the Escherichia coli ATCC 25922, Salmonella Typhimurium ATCC 14028, and Staphylococcus aureus ATCC 25923. The results of PCR amplification revealed that only five strains possess virulence factor genes (gelE,asa1,cyl A,esp). We determined antibiotic resistance, biofilm forming abilities, and hemolytic activity for safety evaluation of strains. CONCLUSIONS: In this large and comprehensive study, we found that only few of Enterococcus strains have promising probiotic potential, among which E. faecalis ES1 and E. faecium EM1 showed the best probiotic properties (are the most promising probiotic candidates).

Effect of cooking temperature on alkaline phosphatase in the production of raw-milk Pecorino cheese.

Alkaline phosphatase (ALP) is a native raw-milk enzyme used in many countries as the standard assay for rapidly validating the milk pasteurization process. Due to the increased restrictions on the production or import of cheeses produced from unpasteurized milk, ALP activity (<10 mU/g) in cheese was measured as a simple and reliable method to check proper milk pasteurization in cheese for both safety inspection and trading controls. In Sicily, the artisanal cheesemaking of the Protected Denomination of Origin (PDO) semi-hard cheeses made with raw sheep milk, includes the cooking of the curd, after whey separation, in a wooden vat under hot Scotta whey (≥80°C), for 3 to 4 h, and finally is left to cool at ambient temperature. Thus, the temperatures adopted during cheesemaking may inactivate the ALP enzyme. To this purpose, the aim of this study was to demonstrate how different temperatures of Scotta whey (35°C [T35], 60°C [T60], 70°C [T70], 80°C [T80], 90°C [T90], and 100°C [T100]) used during the second cooking of Pecorino cheeses after molding for 3 h, influence the ALP activity in fresh and 3-mo aged cheese, both at core and outside. The results highlight that the rate of reduction of ALP was greater with increasing temperature of the second cooking, in particular for T 80°C curd, indicating that the use of Scotta whey >80°C could be a breakpoint able to reduce the ALP activity to values <10 mU/g. Different effects between the core and the outside portions of the experimental cheeses were found, with a decrease in ALP activity more on the outside than in the core portions, in both fresh and 3-mo aged cheeses, for T80, T90, and T100 treatments. Care must be taken in using ALP to control the use of pasteurized milk in the production of PDO cheeses without considering the cheesemaking processes, such as the second cooking, which could be equal to pasteurization, and an adequate interaction of time and temperature can reduce the ALP activity to values comparable with cheeses produced with pasteurized milk.

Evaluation of Polycyclic Aromatic Hydrocarbons in Smoked Cheeses Made in Poland by HPLC Method.

Smoked cheeses are particularly popular among consumers for their flavor and aroma. Of interest, therefore, is the health aspect related to the likelihood of polycyclic aromatic hydrocarbons (PAHs), which are known carcinogens found in smoked products. Thus, the purpose of this study was to evaluate the occurrence of 15 polycyclic aromatic hydrocarbons (PAHs) in smoked and non-smoked cheeses purchased in Poland to monitor their safety. The level of selected PAHs in cheese samples was determined using the HPLC-DAD-FLD method. Most of the cheeses tested met the maximum level of benzo[a]pyrene (2 µg/kg) and the sum of benz[a]anthracene, chrysene, benzo[b]fluoranthene and benzo[a]pyrene (12 µg/kg) established for these products. However, all the cheeses studied in this work had relatively low amounts of the sum of these compounds compared to the information available in the cheese literature, ranging from

Fungal community and physicochemical profiles of ripened cheeses from the Canastra of Minas Gerais, Brazil.

Canastra's Minas artisanal cheese [QMA (Minas artisanal cheese)] is a protected geographical indication traditional food. The influence of fungi on the cheese ripening process is of great importance. This study aimed to apply culture-dependent and -independent methods to determine the mycobiota of QMA produced in the Canastra region, as well as to determine its physicochemical characteristics. Illumina-based amplicon sequencing and matrix-assisted laser desorption ionization time-of-flight mass spectrometry were the culture-independent methods used. The physicochemical analysis results showed that the QMA has a moisture content ranging 18.4-28.2%, fat content ranging 20.5-40%, sodium chloride percentage of approximately 0.9%, and pH ranging 5.2-5.5. The population of fungi ranged between 6.3 and 8 log colony-forming unit/g. Fusarium spp., Geotrichum candidum, Paecilomyces spp., Trichosporon coremiiforme, Candida catenulata, Aspergillus spp., Trichosporon japonicum, Aspergillus oryzae, Kluyveromyces spp., Torulaspora spp., and Debaryomyces spp. were the most prevalent fungi. The methods used to evaluate the mycobiota provide a better understanding of which species are present in the final product and eventually contribute to the characteristics of QMA. Geotrichum candidum and C. catenulata were identified as promising species for future studies on product quality.

Antioxidant capacity and identification of radical scavenging peptides from Crema de Chiapas, Fresco and Cocido cheeses.

Bioactive peptides may positively impact bodily functions. One of these are the antioxidant peptides which are well documented for a wide variety of food matrices, mostly from plant sources. Nevertheless, information of antioxidant milk-derived peptides is still a little-known field. The present study was aimed to evaluating the antioxidant capacity (AC) in vitro of water soluble extracts < 3 kDa (WSE) from three artisanal Mexican cheeses: Crema de Chiapas (CrC), Fresco (FC) and Cocido (CC). This study was carried out for cheeses of different days of storage (0, 5, 10, 15, and 20) at 4 °C. AC was assayed to the respective WSE by 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid) diamoniun salt (ABTS) and oxygen radical absorbance capacity (ORAC) methods and those WSE that showed the most antioxidant capacity from each cheese were analyzed by using RP-HPLC/MS to identify and characterize the novel specific peptides. All the WSE analyzed show antioxidant capacity, especially those from CrC and CC which display the highest AC at 15 days of storage. Regarding to WSE from FC, the AC was constant during storage. Identified structures reveal that these novel peptides possess high content of specific amino acids, mainly proline, valine, leucine and phenylalanine, of which it has already been shown antioxidant properties. This study demonstrate that these artisanal Mexican cheeses are sources of potential antioxidant peptides.

In-Depth Investigation of the Safety of Wooden Shelves Used for Traditional Cheese Ripening.

The main goal of this research was to characterize the bacterial diversity of the wooden boards used for aging traditional Sicilian cheeses and to evaluate whether pathogenic bacteria are associated with these surfaces. Eighteen cheese dairy factories producing three traditional cheese typologies (PDO Pecorino Siciliano, PDO Piacentinu Ennese, and Caciocavallo Palermitano) were selected within the region of Sicily. The wooden shelf surfaces were sampled by a destructive method to detach wood splinters as well as by a nondestructive brushing to collect microbial cells. Scanning electron microscopy showed the presence of almost continuous bacterial formations on the majority of the shelves analyzed. Yeasts and fungal hyphae were also visualized, indicating the complexity of the plank communities. The amplicon library of the 16S rRNA gene V3-V4 region was paired-end sequenced using the Illumina MiSeq system, allowing the identification of 14 phyla, 32 classes, 52 orders, 93 families, and 137 genera. Staphylococcus equorum was identified from all wooden surfaces, with a maximum abundance of 64.75%. Among cheese-surface-ripening bacteria, Brevibacterium and Corynebacterium were detected in almost all samples. Several halophilic (Halomonas, Tetragenococcus halophilus, Chromohalobacter, Salimicrobium, Marinococcus, Salegentibacter, Haererehalobacter, Marinobacter, and Idiomarinaceae) and moderately halophilic (Salinicoccus, Psychrobacter, and Salinisphaera) bacteria were frequently identified. Lactic acid bacteria (LAB) were present at low percentages in the genera Leuconostoc, Lactococcus, Lactobacillus, Pediococcus, and Streptococcus. The levels of viable microorganisms on the wooden shelves ranged between 2.4 and 7.8 log CFU/cm2. In some cases, LAB were counted at very high levels (8.2 log CFU/cm2). Members of the Enterobacteriaceae family were detected in a viable state for only six samples. Coagulase-positive staphylococci, Salmonella spp., and Listeria monocytogenes were not detected. Seventy-five strains belonged to the genera Leuconostoc, Lactococcus, Pediococcus, Enterococcus, Lactobacillus, and Weissella. IMPORTANCE This study provides evidence for the lack of pathogenic bacteria on the wooden shelves used to ripen internal bacterially ripened semihard and hard cheeses produced in Sicily. These three cheeses are not inoculated on their surfaces, and surface ripening is not considered to occur or, at least, does not occur at the same extent as surface-inoculated smear cheeses. Several bacterial groups identified from the wooden shelves are typically associated with smear cheeses, strongly suggesting that PDO Pecorino Siciliano, PDO Piacentinu Ennese, and Caciocavallo Palermitano cheese rind contributes to their final organoleptic profiles.

Microbial profiles of Greek PDO cheeses assessed with amplicon metabarcoding.

Greece is a country possessing many cheese products granted with a PDO (Protected Designation of Origin) certificate, with high exporting activities. In this study, we analyzed six popular cheese PDO products purchased from different industries to assess their microbial communities using amplicon metabarcoding analysis. To this end, using Next Generation Sequencing technology, we sequenced the 16S rRNA gene and the ITS spacer for prokaryotes and fungi, respectively. Alpha diversity indices revealed higher bacterial species richness for some cheeses (Kopanisti, Batzos) and poor for others (Feta, Galotiri). Kopanisti, together with Kalathaki and Anevato, also presented increased species diversity concerning fungal populations. Results showed that lactic acid bacteria (LAB) prevailed the bacterial populations in all samples (Lactococcus, Lactobacillus, Streptococcus, Leuconostoc), whereas for fungi, members of the Saccharomycetaceae, Dipodascaceae and Debaryomycetaceae families prevailed the fungal populations. Several other genera were identified that make up each product's microbiome leading to the creation of the unique organoleptic attributes of Greek PDO cheeses. However, the identified species could not be directly linked to certain cheese types, assuming that starter and adjunct cultures, combined with the raw material used during production greatly impact the microbial communities in cheeses. Our data, produced for the first time for six Greek PDO cheeses, can be exploited in the process of creating a core microbial signature within each cheese type, supporting the Greek brand name and valorizing cheese products.

Directed Recovery and Molecular Characterization of Antibiotic Resistance Plasmids from Cheese Bacteria.

Resistance to antimicrobials is a growing problem of worldwide concern. Plasmids are thought to be major drivers of antibiotic resistance spread. The present work reports a simple way to recover replicative plasmids conferring antibiotic resistance from the bacteria in cheese. Purified plasmid DNA from colonies grown in the presence of tetracycline and erythromycin was introduced into plasmid-free strains of Lactococcus lactis, Lactiplantibacillus plantarum and Lacticaseibacillus casei. Following antibiotic selection, the plasmids from resistant transformants were isolated, analyzed by restriction enzyme digestion, and sequenced. Seven patterns were obtained for the tetracycline-resistant colonies, five from L. lactis, and one each from the lactobacilli strains, as well as a single digestion profile for the erythromycin-resistant transformants obtained in L. lactis. Sequence analysis respectively identified tet(S) and ermB in the tetracycline- and erythromycin-resistance plasmids from L. lactis. No dedicated resistance genes were detected in plasmids conferring tetracycline resistance to L. casei and L. plantarum. The present results highlight the usefulness of the proposed methodology for isolating functional plasmids that confer antibiotic resistance to LAB species, widen our knowledge of antibiotic resistance in the bacteria that inhabit cheese, and emphasize the leading role of plasmids in the spread of resistance genes via the food chain.

Content and Nutritional Evaluation of Zinc in PDO and Traditional Italian Cheeses.

Zinc is an essential mineral which plays a key role in several important biological processes in the human body. The determination of its level in food matrices can contribute to the food quality characterization and to the adequacy of the diet. Animal food products generally have a higher zinc content compared to vegetables. Among them, dairy products consumption can provide a great contribution to the zinc reference intakes. In this study, different Italian cheeses (38 Protected Denomination of Origin and 9 Traditional) were evaluated for their zinc content. Cow cheeses generally showed the highest zinc content (1.83-7.75 mg/100 g cheese), followed by sheep cheeses (1.34-3.69 mg/100 g), and cheeses from mixed milk (0.39-4.54 mg/100 g). The only cheese from buffalo milk (Mozzarella di Bufala Campana PDO) showed a zinc content of 2.14 mg/100 g. The great variability in the zinc content observed among the samples is the result of the influence of several factors, such as the feeding system, the species (cow, sheep, goat, and buffalo), and the cheese-making. Most of the samples resulted in a great contribution (>10%) to the zinc Daily Reference Intake set by EU (10 mg/day), with only two samples contributing to less than 4%.

The Portuguese Serrana goat breed: a review.

Goats were among the first animals to be domesticated over 10,000 years ago and are part of human societies since the beginning of agriculture. Goats play a major role both in commercial farming systems and in subsistence agriculture systems, particularly in tropical, subtropical and Mediterranean regions where they are crucial for the supply of meat, milk, fibre and dung. This review concerns the Serrana breed, the most important and numerous indigenous goat breed from Portugal that was furthermore exported to other regions of the world, notably South America during the Portuguese colonization. Herein, we describe the origin and history of the breed as well as the productive performance and most common production systems. Finally, we address the local and traditional PDO (protected denomination of origin) and PGI (protected geographical indication) that are produced from these animals.

Identification of bacterial hazards in the production of artisan fresh cheese in Cuba.

Artisan fresh cheese producing farms from six provinces of Cuba were studied to identify the presence of bacterial hazards and the results are presented in this research communication. The bacterial hazards identified in milk and cheese respectively were: Listeria spp. (9.5 and 18.9%), Bacillus cereus (23.2 and 24.2%), Escherichia coli O157 (12.6 and 13.7%), Salmonella spp. (10.5 and 17.9%), and Staphylococcus aureus (29.5 and 51.6%). Listeria monocytogenes was not detected. Nine Salmonella serotypes corresponding to Salmonella enterica subsp. enterica and Salmonella enterica subsp. arizonae were isolated, whereas Salmonella Anatum was present most often. Biofilm formation by the isolated species and enterotoxin production by S. aureus strains demonstrated the pathogenic potential of the identified bacterial hazards. Results proved the presence of bacterial hazards in the raw milk and cheeses analyzed, so that good manufacturing practices must be accomplished throughout the entire production process in order to avoid the occurrence of foodborne diseases in the population.

Colorants in cheese manufacture: Production, chemistry, interactions, and regulation.

Colored Cheddar cheeses are prepared by adding an aqueous annatto extract (norbixin) to cheese milk; however, a considerable proportion (∼20%) of such colorant is transferred to whey, which can limit the end use applications of whey products. Different geographical regions have adopted various strategies for handling whey derived from colored cheeses production. For example, in the United States, whey products are treated with oxidizing agents such as hydrogen peroxide and benzoyl peroxide to obtain white and colorless spray-dried products; however, chemical bleaching of whey is prohibited in Europe and China. Fundamental studies have focused on understanding the interactions between colorants molecules and various components of cheese. In addition, the selective delivery of colorants to the cheese curd through approaches such as encapsulated norbixin and microcapsules of bixin or use of alternative colorants, including fat-soluble/emulsified versions of annatto or beta-carotene, has been studied. This review provides a critical analysis of pertinent scientific and patent literature pertaining to colorant delivery in cheese and various types of colorant products on the market for cheese manufacture, and also considers interactions between colorant molecules and cheese components; various strategies for elimination of color transfer to whey during cheese manufacture are also discussed.

Chemical-nutritional characteristics and aromatic profile of milk and related dairy products obtained from goats fed with extruded linseed.

OBJECTIVE: This study aimed to evaluate the effect of dietary integration with extruded linseed (EL) on fatty acid (FA) and aromatic profile of goat cheese after 60 (T60) days of ripening. METHODS: Thirty goats were divided in two groups. The control group (CG) was fed with conventional diet, whereas the experimental group (EL+) was fed with conventional diet supplemented with 10% of EL. Milk samples were collected on 30 and 60 days of trial to determinate chemical-nutritional composition and FA profile. At the end of experiment, six cheese-making session (3 for each group) were carried out using a pooled milk sample obtained from the 15 goat of each group. At 60 days of ripening, cheeses were analyzed for chemical-nutritional composition, FA and aromatic profile. RESULTS: An increase in the milk production, protein, fat and lactose were evidenced in the EL+ goats. Conversely, a reduction of somatic cells was observed in the EL+ compared with the CG. However, no variation was observed for urea and casein levels content in milk samples, and no changes in protein and lipid content were found for cheeses in the two experimental groups. Dietary supplementation with EL modified the FA profile of milk. There was a decrease in saturated fatty acids (SFA) and an increase in polyunsaturated fatty acids (PUFA). Chemical composition of T60 cheese did not differ between the two groups but a different FA profile was observed. In particular, in T60 cheese obtained from EL+ milk, an increase in short-chain FA (SCFA) and a decrease in medium and long-chain FA (MCFA and LCFA) were observed. The EL diet led to cheeses with butanoic acid 2 time higher compared to CG cheeses. Moreover, a greater presence of aldehyde compounds and alcohols were observed in the cheeses of experimental group. CONCLUSION: The present study pointed out that EL supplementation may improve the chemical and physical qualities of goat milk and cheeses.

Large-scale mapping of microbial diversity in artisanal Brazilian cheeses.

Brazilian artisanal cheeses are characterized by the use of raw milk and in some cases, natural starter cultures, known as "pingo", as well as following simple and traditional manufacturing technology. In this study, a large-scale screening of the microbial ecology of 11 different types of artisanal cheeses produced in five geographical areas of Brazil was performed. Besides, the specific origin-related microbial signatures were identified. Clear geography- and technology-based differences in the microbiota were observed. Lactic acid bacteria dominated in all cheeses although Enterobacteriaceae and Staphylococcus also occurred in North, Northeast and Central cheeses. Differences in the lactic acid bacteria patterns were also highlighted: Streptococcus, Leuconostoc, Lactococcus and Lactobacillus were differently combined in terms of relative abundance according to product type and region of production. This study provides a comprehensive, unprecedented microbiological mapping of Brazilian cheeses, highlighting the impact of geographical origin and mode of production on microbial diversity. The results obtained will help to plan an evaluation of microbial contamination sources that will need to be studied for the improvement of cheese quality and safety.

Bacterial microbiota of Kazakhstan cheese revealed by single molecule real time (SMRT) sequencing and its comparison with Belgian, Kalmykian and Italian artisanal cheeses.

BACKGROUND: In Kazakhstan, traditional artisanal cheeses have a long history and are widely consumed. The unique characteristics of local artisanal cheeses are almost completely preserved. However, their microbial communities have rarely been reported. The current study firstly generated the Single Molecule, Real-Time (SMRT) sequencing bacterial diversity profiles of 6 traditional artisanal cheese samples of Kazakhstan origin, followed by comparatively analyzed the microbiota composition between the current dataset and those from cheeses originated from Belgium, Russian Republic of Kalmykia (Kalmykia) and Italy. RESULTS: Across the Kazakhstan cheese samples, a total of 238 bacterial species belonging to 14 phyla and 140 genera were identified. Lactococcus lactis (28.93%), Lactobacillus helveticus (26.43%), Streptococcus thermophilus (12.18%) and Lactobacillus delbrueckii (12.15%) were the dominant bacterial species for these samples. To further evaluate the cheese bacterial diversity of Kazakhstan cheeses in comparison with those from other geographic origins, 16S rRNA datasets of 36 artisanal cheeses from Belgium, Russian Republic of Kalmykia (Kalmykia) and Italy were retrieved from public databases. The cheese bacterial microbiota communities were largely different across sample origins. By principal coordinate analysis (PCoA) and multivariate analysis of variance (MANOVA), the structure of the Kazakhstan artisanal cheese samples was found to be different from those of the other geographic origins. Furthermore, the redundancy analysis (RDA) identified 16 bacterial OTUs as the key variables responsible for such microbiota structural difference. CONCLUSION: Our results together suggest that the diversity of bacterial communities in different groups is stratified by geographic region. This study does not only provide novel information on the bacterial microbiota of traditional artisanal cheese of Kazakhstan at species level, but also interesting insights into the bacterial diversity of artisanal cheeses of various geographical origins.

Enterotoxin production and antimicrobial susceptibility in Staphylococci isolated from traditional raw milk cheeses in Serbia.

This study was undertaken to determine the prevalence of coagulase positive staphylococci (CPS) by examining a total of 71 raw milk cheeses. Additionally, enterotoxigenicity, antimicrobial susceptibility and the presence of mecA and mecC genes in the staphylococcal isolates were investigated. The isolation and enumeration procedure of CPS followed the International Organization for Standardization (ISO) standard. The presumptive staphylococci were identified by matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) using the VITEK MS system. VIDAS® Staph enterotoxin II assay was used for the detection of classical enterotoxins. Antimicrobial susceptibility testing (AST) was accomplished performing the disk diffusion method. All suspected methicillin resistant staphylococci were investigated for the presence of mecA and mecC genes by PCR assay. A high prevalence (87.32%) of CPS was detected in the cheeses at contamination levels up to 5.58 log CFU g-1. Among 47 staphylococcal isolates screened for enterotoxin production, only one isolate, identified as S. hyicus, was confirmed as being enterotoxigenic. Resistance to penicillin (63.70%) was the most common resistance among the tested Staphylococcus aureus isolates. The dominant phenotypic resistance patterns in coagulase negative staphylococci (CNS) were resistance to ofloxacin and fusidic acid. All CNS isolates were susceptible to the clinically important antibiotics clindamycin, chloramphenicol, gentamicin, linezolid, rifampicin and trimethoprim-sulfamethoxazole. The mecA and mecC genes were not detected. To the best of our knowledge, this is the first study concerning evaluation of the presence of methicillin resistant staphylococci (MRS) in dairy products in Serbia.

Control of Shigatoxin-producing Escherichia coli in cheese by dairy bacterial strains.

Bio-preservation could be a valuable way to control Shigatoxin-producing Escherichia coli (STEC) in cheese. To this end, 41 strains were screened for their inhibitory potential on model cheese curd and on pasteurized and raw milk uncooked pressed cheeses. Strains of Lactococcus lactis, Lactococcus garvieae, Leuconostoc pseudomesenteroides, Leuconostoc citreum, Lactobacillus sp, Carnobacterium mobile, Enterococcus faecalis, Enterococcus faecium, Macrococcus caseolyticus and Hafnia alvei reduced STEC O26:H11 counts by 1.4-2.5 log cfu g(-1) and to a lesser extent STEC O157:H7 counts in pasteurized milk cheeses. Some strains can act in synergy to inhibit STEC in raw milk uncooked pressed cheeses. Inhibitory associations had no adverse effect on the sensory characteristics of these cheeses. The association of H. alvei, Lactobacillus plantarum and Lc. lactis was the most inhibitory: after inoculation of this consortium into milk, STEC O26:H11 and O157:H7, inoculated at 2 log cfu ml(-1), were reduced by up to 3 log cfu g(-1) in ripened cheese. Inhibition in cheese cannot be predicted from H2O2 production in BHI medium, decreased pH or milk reduction. It is not clear what role the rapid decrease in pH during the first 6 h may play in the inhibition. Further studies will be needed to determine the nature of the inhibition.

Characterization of yeasts isolated from artisanal short-ripened cows' cheeses produced in Galicia (NW Spain).

A total of 143 presumptive yeast isolates were obtained from the predominant microflora of 21 short-ripened starter-free raw cow's milk cheeses made in Galicia (NW Spain), and the following 68 isolates were identified by both genotyping and sequencing methods: Yarrowia lipolytica (21 isolates), Kluyveromyces lactis (18), Debaryomyces hansenii (11), Pichia guilliermondii (11), Pichia fermentans (4) and Saccharomyces cerevisiae (3). Of these, Y. lipolytica and K. lactis displayed the strongest extracellular proteolytic activity on skim milk agar, and none of the D. hansenii isolates showed any activity on this medium. Y. lipolytica also displayed the highest lipolytic activity on Tween 80 and on tributyrin. This species, which was characterized by production of butanoic acid, free fatty acid esters and sulfur compounds in pasteurized whole milk, was responsible for rancid and cheesy flavors. K. lactis mainly produced acetaldehyde, ethanol, branched chain aldehydes and alcohols, and acetic acid esters, which were responsible for alcoholic, fruity and acetic notes. The volatile profiles of D. hansenii were rather limited and characterized by high levels of methyl ketones. Most of the yeast isolates were described as tryptamine producers, although low concentrations of histamine were produced by five Y. lipolytica and two P. fermentans isolates. We conclude that selected Y. lipolytica strains could be used as adjunct cultures in the manufacture of Arzúa-Ulloa and Tetilla cheeses, and selected K. lactis strains could be used as co-starters in the manufacture of acid curd Cebreiro cheese, thus contributing to the sensory quality and typicality of the cheeses.