Influence of an autochthonous starter culture and a commercial starter on the characteristics of Tenerife pasteurised goats’ milk cheese

Two batches of Tenerife cheese were produced from pasteurised goats’ milk, one with a commercial starter and another with an autochthonous starter consisting of three selected strains of lactic acid bacteria isolated from artisanal Tenerife cheese. Influence of starter on the characteristics of the cheeses was evaluated. Owing to the composition of the starters, lactobacilli and leuconostocs varied significantly between both cheeses. However, the type of starter did not significantly affect the physicochemical and proteolytic properties of the cheeses. In general, cheeses made with autochthonous starter received higher scores for sensory attributes, especially for aroma, than those made with commercial starter.     

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.     

Functionality of enterococci in dairy products

Enterococci have important implications in the dairy industry. They occur as nonstarter lactic acid bacteria (NSLAB) in a variety of cheeses, especially artisan cheeses produced in southern Europe from raw or pasteurised milk, and in natural milk or whey starter cultures. They play an acknowledged role in the development of sensory characteristics during ripening of many cheeses and have been also used as components of cheese starter cultures. The positive influence of enterococci on cheese seems due to specific biochemical traits such as lipolytic activity, citrate utilisation, and production of aromatic volatile compounds. Some enterococci of dairy origin have also been reported to produce bacteriocins (enterocins) inhibitory against food spoilage or pathogenic bacteria, such as Listeria monocytogenes, Staphylococcus aureus, Vibrio cholerae, Clostridium spp., and Bacillus spp. The technological application of enterocins, shown to be produced during cheese manufacture, led to propose enterococci as adjunct starter or protective cultures in cheeses. There is evidence that enterococci, either added as adjunct starters or present as nonstarter NSLAB, could find potential application in the processing of some fermented dairy products. Literature suggest that the complex biochemical and ecological phenomena explaining the technological functionality of the enterococci in dairy products, are still to be fully understood. Clearly, the clinical research on enterococci underlines also that the safety of dairy products containing enterococci is an issue that the industry must carefully address before proceeding to their application.     

Angiotensin-converting enzyme inhibitory activity of milk fermented by wild and industrial Lactococcus lactis strains

Angiotensin I-converting enzyme inhibitory (ACEI) activity was evaluated and compared in <3 KDa water-soluble extracts (WSE) isolated from milk fermented by wild and commercial starter culture Lactococcus lactis strains after 48 h of incubation. The highest ACEI activities were found in WSE from milk inoculated with wild L. lactis strains isolated from artisanal dairy products and commercial starter cultures. On the other hand, the lowest ACEI activities were found in WSE from milk inoculated with wild strains isolated from vegetables. Moreover, the IC₅₀ values (concentration that inhibits 50% activity) of WSE from artisanal dairy products were the lowest, indicating that these fractions were the most effective in inhibiting 50% of ACE activity. In fact, a strain isolated from artisanal cheese presented the lowest IC₅₀ (13 μg/mL). Thus, it appears that wild L. lactis strains isolated from artisanal dairy products and commercial starter cultures showed good potential for the production of fermented dairy products with ACEI properties.     

A high-throughput cheese manufacturing model for effective cheese starter culture screening

Cheese making is a process in which enzymatic coagulation of milk is followed by protein separation, carbohydrate removal, and an extended bacterial fermentation. The number of variables in this complex process that influence cheese quality is so large that the developments of new manufacturing protocols are cumbersome. To reduce screening costs, several models have been developed to miniaturize the cheese manufacturing process. However, these models are not able to accommodate the throughputs required for systematic screening programs. Here, we describe a protocol that allows the parallel manufacturing of approximately 600 cheeses in individual cheese vats each with individual process specifications. Protocols for the production of miniaturized Gouda- and Cheddar-type cheeses have been developed. Starting with as little as 1.7 mL of milk, miniature cheeses of about 170 mg can be produced and they closely resemble conventionally produced cheese in terms of acidification profiles, moisture and salt contents, proteolysis, flavor profiles, and microstructure. Flavor profiling of miniature cheeses manufactured with and without mixed-strain adjunct starter cultures allowed the distinguishing of the different cheeses. Moreover, single-strain adjunct starter cultures engineered to overexpress important flavor-related enzymes revealed effects similar to those described in industrial cheese. Benchmarking against industrial cheese produced from the same raw materials established a good correlation between their proteolytic degradation products and their flavor profiles. These miniature cheeses, referred to as microcheeses, open new possibilities to study many aspects of cheese production, which will not only accelerate product development but also allow a more systematic approach to investigate the complex biochemistry and microbiology of cheese making.     

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.     

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

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

Proteolysis on Reggianito Argentino cheeses manufactured with natural whey cultures and selected strains of Lactobacillus helveticus

Reggianito Argentino cheese is traditionally manufactured with whey starter cultures that provide typical and intense flavor but can cause poor quality standardization. In this study, the influence of natural and selected starters on Reggianito Argentino cheese proteolysis was investigated. Cheeses were manufactured with three strains of Lactobacillus helveticus (SF133, SF138 and SF209) cultured individually in sterile whey and used as single or mixed starters. Control cheeses were made with natural whey starter culture. Cheeses were analyzed to determine gross composition, as well as total thermophilic lactic flora. Proteolysis was assessed by N fractions, electrophoresis and liquid chromatography. Gross composition of the cheeses did not significantly differ, while viable starter cell counts were lower for cheeses made with strain SF209 alone or combined with other strains. Soluble N at pH 4.6 was the same for cheeses made with natural or selected starters, but soluble N in 12% trichloroacetic acid and 2.5% phosphotungstic acid was significantly higher in cheeses made with starters containing strain SF209. Nitrogen fractions results indicated that natural whey starter cultures could be replaced by several starters composed of the selected strains without significant changes to proteolysis patterns. Starter cultures prepared only with SF209 or with the three selected L. helveticus strains produced cheese products with significantly more proteolysis than control cheeses. Chromatographic profiles analyzed by principal components showed that three main peaks on chromatograms, presumptively identified as Tyr, Phe, and Trp, explained most of variability. Principal component scores indicated that cheese samples were grouped by ripening time, which was confirmed by linear discriminant analysis. On the contrary, samples did not cluster by Lactobacillus strain or type of starter.     

Brazilian Artisanal Cheeses: An Overview of their Characteristics,Main Types and Regulatory Aspects

A large variety of cheeses exist in Brazil, reflecting historical and cultural aspects. Brazilian artisanal cheeses present differences in the processing, ripening time (when applied), type of milk used, texture, size, shape, color, moisture content, flavor, the use or not of starter cultures, among others. This review describes the main artisanal cheeses produced in Brazil, focusing on general and particular characteristics associated with their making process and geographical identity. Overall, the high variability of the physicochemical data and deficiency of information on sensorial properties of Brazilian artisanal cheeses were noticed. On the other hand, culture‐dependent methods were mostly used to expand the knowledge into the microbiology of these cheeses, whereas their microbial diversity has been recently discovered through the use of 16S rRNA gene sequencing‐based methods. The certification of a geographical indication for Brazilian artisanal cheeses may encompass an essential milestone for adding value to these products. Regardless of their significance in the diet, culture, and economy of producing regions, taken together, the reviewed literature discloses the need of insightful studies to generate scientific data to support the expansion of the market, while ensuring the protection of historic aspects related to the production of Brazilian artisanal cheeses.     

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.     

Major advances in concentrated and dry milk products, cheese, and milk fat-based spreads

Advances in dairy foods and dairy foods processing since 1981 have influenced consumers and processors of dairy products. Consumer benefits include dairy products with enhanced nutrition and product functionality for specific applications. Processors convert raw milk to finished product with improved efficiencies and have developed processing technologies to improve traditional products and to introduce new products for expanding the dairy foods market. Membrane processing evolved from a laboratory technique to a major industrial process for milk and whey processing. Ultra-filtration and reverse osmosis have been used extensively in fractionation of milk and whey components. Advances in cheese manufacturing methods have included mechanization of the making process. Membrane processing has allowed uniform composition of the cheese milk and starter cultures have become more predictable. Cheese vats have become larger and enclosed as well as computer controlled. Researchers have learned to control many of the functional properties of cheese by understanding the role of fat and calcium distribution, as bound or unbound, in the cheese matrix. Processed cheese (cheese, foods, spreads, and products) maintain their importance in the industry as many product types can be produced to meet market needs and provide stable products for an extended shelf life. Cheese delivers concentrated nutrients of milk and bio-active peptides to consumers. The technologies for the production of concentrated and dried milk and whey products have not changed greatly in the last 25 yr. The size and efficiencies of the equipment have increased. Use of reverse osmosis in place of vacuum condensing has been proposed. Modifying the fatty acid composition of milkfat to alter the nutritional and functional properties of dairy spread has been a focus of research in the last 2 decades. Conjugated linoleic acid, which can be increased in milkfat by alteration of the cow's diet, has been reported to have anticancer, anti-atherogenic, antidiabetic, and antiobesity effects for human health. Separating milk fat into fractions has been accomplished to provide specific fractions to improve butter spreadability, modulate chocolate meltability, and provide texture for low-fat cheeses.     

Artisanal and experimental Pecorino Siciliano cheese: microbial dynamics during manufacture assessed by culturing and PCR-DGGE analyses

Traditional artisanal Pecorino Siciliano (PS) cheeses, and two experimental PS cheeses were manufactured using either raw or pasteurised ewes' milk with the addition of starter cultures. The bacterial diversity and dynamics of the different cheese types were evaluated both by culturing and characterisation of isolates, and a culture-independent approach based on the 16S ribosomal RNA (rRNA) gene. Following cultivation, artisanal and experimental cheese types showed similar microbial counts, and isolates belonging to Lactococcus lactis, Streptococcus thermophilus, Enterococcus faecalis and Leuconostoc mesenteroides were identified by phenotypic characterisation and comparison of the restriction fragment length polymorphism (RFLP) of the 16S rRNA gene to that of reference species. The culture-independent fingerprinting technique PCR and denaturing gradient gel electrophoresis (DGGE) of V6 to V8 regions of the 16S rRNA gene of samples taken during artisanal PS cheese manufacture, from raw milk to the ripened cheese, indicated relevant shifts in the microbial community structure. The dominance of Streptococcus bovis and Lactococcus lactis species in the traditional artisanal PS was revealed by 16S rRNA gene sequencing. Comparison of DGGE profiles of samples from milk to ripened cheese, derived from artisanal procedure and the two experimental PS cheeses during production showed similar trends with the presence of intense bands in common. Nevertheless, the profiles of several artisanal cheeses from different farms appeared more diverse, and these additional species are probably responsible for the generally superior flavour and aroma development of traditional PS cheese.     

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.     

Rational selection of Leuconostoc strains for mixed starters based on the physiological biodiversity found in raw milk fermentations

The technological abilities of eleven wild Leuconostoc strains isolated from artisanal Afuega'l Pitu cheese, a homemade acid-coagulated cheese from raw milk from Asturias (northern Spain), were studied in order to test their potential application as components of dairy starters. Metabolic activity, production of flavour compounds, resistance to NaCl, acid, nisin and freezing, as well as genetic biodiversity were investigated. Marked differences among all tested strains reflected the existing biodiversity in naturally fermented products. After evaluation of their performance, strains IPLA567 and IPLA979 revealed the best properties to be used use in mixed dairy starter cultures. These two strains were able to ferment lactose and galactose and produced larger amounts of lactic acid than the others. Moreover, they showed high tolerance levels to NaCl, acid and nisin, although their resistance to freezing was slightly lower than the other strains. This study evidences the fact that natural environments can be considered as a proper source of new strains, which may help the dairy industry to widen their fermented products range.     

Selection, application and monitoring of Lactobacillus paracasei strains as adjunct cultures in the production of Gouda-type cheeses

Raw milk cheeses have more intense flavours than cheeses made from pasteurized milk and harbour strains with potential adjunct properties. Two Lactobacillus paracasei strains, R-40926 and R-40937, were selected as potential adjunct cultures from a total of 734 isolates from good quality artisan raw milk Gouda-type cheeses on the basis of their prevalence in different cheese types and/or over several production batches, safety and technological parameters. Conventional culturing, isolation and identification and a combined PCR-DGGE approach using total cheese DNA extracts and DNA extracts obtained from culturable fractions were employed to monitor viability of the introduced adjuncts and their effect on the cheese microbiota. The control cheese made without adjuncts was dominated by members of the starter, i.e. Lactococcus lactis and Leuconostoc pseudomesenteroides. In the cheeses containing either R-40926 or R-40937, the respective adjuncts increased in number as ripening progressed indicating that both strains are well adapted to the cheese environment and can survive in a competitive environment in the presence of a commercial starter culture. Principal component analysis of cheese volatiles determined by steam distillation-extraction and gas chromatography-mass spectrometry could differentiate cheeses made with different concentrations of adjunct R-40926 from the control cheese, and these differences could be correlated to the proteolytic and lipolytic properties of this strain. Collectively, results from microbiological and metabolic analyses indicate that the screening procedure followed throughout this study was successful in delivering potential adjunct candidates to enrich or extend the flavour palette of artisan Gouda-type cheeses under more controlled conditions.     

Influence of pasteurised milk, raw milk and different ripening cultures on biogenic amine concentrations in semi-soft cheeses during ripening

In semi-soft cheeses, produced with pasteurised milk, raw milk and different starter cultures, the concentrations of cadaverine, histamine, phenylethylamine, putrescine and tyramine were investigated. The cultures (pasteurised milk cultures, raw milk cultures and starter cultures) strongly influenced the biogenic amine concentrations in the cheeses ripened for 5 months. Two cheeses made with identical pasteurised milk, but different ripening cultures, differed greatly in their total biogenic amine concentrations (51 vs 371?mg/kg). In general, the biogenic amine concentrations increased markedly between month 2 and month 3 of cheese ripening. The high content of enterococci and Enterobacteriaceae yielded the biogenic amine concentrations. In contrast, Lactobacilli did not seem to be important. However, unspecified bacteria have to be considered, since cheeses with comparable microbiological profiles differed enormously in their biogenic amine concentrations. Semi-soft cheeses produced from pasteurised milk showed remarkably lower total biogenic amine concentrations compared to semi-soft cheeses produced from raw milk (51–1096?mg/kg vs 1011–3133?mg/kg, depending also on the ripening cultures). The highest total biogenic amine concentration (4817?mg/kg) was detected in a cheese produced from raw milk that had been stored for 36?h. In this cheese, the concentrations of cadaverine, phenylethylamine, putrescine and tyramine were higher than in all other cheeses. The highest histamine concentration was found to be in another raw milk cheese (573?mg/kg).     

Comparison of Fascal cheese produced with natural,commercial or autochthonous cultures

Lactic acid bacteria were isolated from raw ovine milk and Fascal cheese and used as two alternative inoculants for production of this Brazilian cheese. Microbiological counts and moisture of these cheeses were comparable to those observed in cheese made with a commercial starter. All cheeses showed absence of Salmonella spp., tolerable counts of coliforms, but coagulase‐positive staphylococci counts exceeded the standard values. Cheeses produced with Lactobacillus plantarum LCN 28 and Lactobacillus rhamnosus LCN 43 showed better results than those observed for the other experimental combinations. Autochthonous cultures could be beneficial to the manufacture of high quality typical raw milk cheese.     

Selected lactic acid bacteria as a hurdle to the microbial spoilage of cheese: Application on a traditional raw ewes' milk cheese

To evaluate the efficacy of lactic acid bacteria (LAB) to improve the hygienic safety of a traditional raw milk cheese, the raw ewes' milk protected denomination of origin (PDO) Pecorino Siciliano cheese was used as a model system. Different Pecorino Siciliano curds and cheeses were used as sources of autochthonous LAB subsequently used as starter and non-starter LAB. These were screened for their acidification capacity and autolysis. Starter LAB showing the best performance were genotypically differentiated and identified: two strains of Lactococcus lactis subsp. lactis were selected. From the non-starter LAB, Enterococcus faecalis, Lactococcus garvieae and Streptococcus macedonicus strains were selected. The five cultures were used in individual or dual inocula to produce experimental cheeses in a dairy factory for which production was characterised by high numbers of undesirable bacteria. At 5-month of ripening, the experimental cheeses produced with LAB were characterised by undetectable levels of enterobacteria and pseudomonads and the typical sensory attributes.     

Graduate Student Literature Review: Enterotoxigenic potential and antimicrobial resistance of staphylococci from Brazilian artisanal raw milk cheeses

More than 30 types of artisanal cheeses are known in Brazil; however, microorganisms, such as Staphylococcus spp., can contaminate raw milk cheeses through different sources, from milking to processing. Staphylococcal food poisoning results from the consumption of food in which coagulase-positive staphylococci, mostly Staphylococcus aureus, have developed and produced enterotoxins. In addition, an emerging public health concern is the increasing antimicrobial resistance of some Staphylococcus strains. Furthermore, the ability of Staphylococcus spp. in sharing antibiotic resistance-related genes with other bacteria increases this problem. In light of these observations, this review aims to discuss the presence of, enterotoxins of, and antibiotic-resistant of Staphylococcus spp. in Brazilian artisanal cheese produced with raw milk.     

Influence of microfiltration and adjunct culture on quality of Domiati cheese

The effects of microfiltration and pasteurization processes on proteolysis, lipolysis, and flavor development in Domiati cheese during 2 mo of pickling were studied. Cultures of starter lactic acid bacteria isolated from Egyptian dairy products were evaluated in experimental Domiati cheese for flavor development capabilities. In the first trial, raw skim milk was microfiltered and then the protein:fat ratio was standardized using pasteurized cream. Pasteurized milk with same protein:fat ratio was also used in the second trial. The chemical composition of cheeses seemed to be affected by milk treatment—microfiltration or pasteurization—rather than by the culture types. The moisture content was higher and the pH was lower in pasteurized milk cheeses than in microfiltered milk cheeses at d 1 of manufacture. Chemical composition of experimental cheeses was within the legal limits for Domiati cheese in Egypt. Proteolysis and lipolysis during cheese pickling were lower in microfiltered milk cheeses compared with pasteurized milk cheeses. Highly significant variations in free amino acids, free fatty acids, and sensory evaluation were found among the cultures used in Domiati cheesemaking. The cheese made using adjunct culture containing Lactobacillus delbrueckii ssp. lactis, Lactobacillus paracasei ssp. paracasei, Lactobacillus casei, Lactobacillus plantarum, and Enterococcus faecium received high scores in flavor acceptability. Cheeses made from microfiltered milk received a higher score in body and texture compared with cheeses made from pasteurized milk.