Innovative Caciocavallo cheeses made from a mixture of cow milk with ewe or goat milk

This study assessed and compared the physicochemical, microbiological, and sensorial characteristics of Caciocavallo cheeses, made from cow milk and a mixture of cow with ewe or goat milk, during ripening. Different cheese-making trials were carried out on an industrial scale following the standard procedure of pasta filata cheeses, with some modifications. The percentage of the different added milk to cow milk influenced compositional and nutritional characteristics of the innovative products, leading to a satisfactory microbiological and sensorial quality.     

Evolution of free amino acids during ripening of Caciocavallo cheeses made with different milks

The evolution of free amino acids (FAA) in Caciocavallo cheeses, made with cow milk (CC) and cow milk mixed with ewe (CE) and goat (CG) milk, was studied throughout ripening. In all Caciocavallo cheeses produced, the total free amino acid (TFAA) content increased during ripening. In general, the highest TFAA content was found in cow cheeses, and the lowest in CG cheeses, whereas CE cheeses ranged over an intermediate level. In all the analyzed samples, during ripening, the content of the individual FAA increased with the exception of arginine. Tyrosine and histidine were found only in CE samples from the middle to the end of ripening. The major FAA found throughout the whole ripening period, in all types of cheese, were leucine, phenylalanine, lysine, valine, asparagine, γ-aminobutyric acid, and ornithine. The TFAA and several AA showed significant differences in ripening time, whereas tyrosine and histidine showed significant differences in kinds of milk.     

Effect of nisin‐producing Lactococcus lactis starter cultures on the inhibition of two pathogens in ripened cheeses

The effect of Lactococcus lactis nisin‐producing strains, isolated from Italian fermented foods, on the survival of two foodborne pathogens namely Listeria monocytogenes and Staphylococcus aureus was investigated in experimental cheese production. One of the three Lactobacillus lactis nisin innoculated as starters, Lactobacillus lactis 41FL1 lowered S. aureus count by 1.73 log colony‐forming units (cfu)/g within the first 3 days, reaching the highest reduction, 3.54 log cfu/g, by the end of ripening period of 60 days. There was no effect on L. monocytogenes. The application of L. lactis 41FL1 as bioprotective culture in controlling S. aureus shows considerable promise.     

Robust PCR‐based method for quantification of bovine milk in cheeses made from caprine and ovine milk

To prevent fraud and enhance quality assurance, credible analysis of dairy products is crucial. A common problem is the addition of cheaper bovine milk to caprine and/or ovine dairy products and when not declared addition of bovine milk constitutes fraud. The aim was to develop a rapid, robust and sensitive method for the identification of adulteration of caprine and/or ovine cheeses with bovine milk. New quantitative real‐time polymerase (qPCR) assays were designed for the specific determination of bovine DNA (Cow1) and bovine, caprine and ovine DNA (BoCaOv). These were applied to 17 samples of caprine cheese and 24 of ovine cheese. Results showed that 17% (7/41) of these cheeses contained >5% bovine milk. As bovine milk was not declared as an ingredient in any of the samples, this represents adulteration. Other cheeses that contained detectable bovine milk at ≤5% (22%; 5/41) might pose a health risk to people allergic to bovine milk.     

A survey of non-starter lactic acid bacteria in traditional cheeses: Culture dependent identification and survival to simulated gastrointestinal transit

Cultivable NSLAB in traditional Pasta filata and ewes' milk cheeses were studied by both PCR-DGGE of cells from Rogosa agar and by isolation and molecular identification after a simulated gastric juice (SGJ) treatment of the cheese. Two to six species were retrieved from each sample. The majority of isolates were identified as Lactobacillus paracasei, Lactobacillus fermentum, Lactobacillus rhamnosus, Enterococcus or Pediococcus. Bile tolerance and bile salt hydrolase (BSH) activity were tested on 88 strains: 64% were able to grow with ≥0.15% bile and 40% were BSH positive. The effect of simulated digestion was tested on 15 strains. Inactivation ranged from 0.15 to 2.93 log cycles; most of the lethality was associated with pancreatic juice treatment. Although SGJ treatment alone may not provide a correct estimate of tolerance to gastrointestinal transit, it allowed selection of strains with a high tolerance to gastric juice, which may be tested as probiotic candidates.     

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.     

Angiotensin‐converting enzyme‐inhibitory and antithrombotic activities of soluble peptide extracts from buffalo and cow milk Cheddar cheeses

The aim of the study was to evaluate potential role of a water‐soluble peptide (WSP) extracts derived from buffalo and cow milk Cheddar cheeses with special reference to their antihypertension and antithrombotic activities. The WSP fractions collected at different stages of ripening were tested to assess their degree of proteolysis, their peptides were profiled by RP‐HPLC and in vitro assays for potential bioactivity were conducted. The peptide peak development was observed with slight differences in peaks number, area and height. Both angiotensin‐converting enzyme‐inhibitory and antithrombotic activities increased progressively during ripening. In comparison, the highest activities were observed in peptide extracts obtained from buffalo milk Cheddar cheese, in a dose‐dependent fashion.     

Effects of salts of organic acids on the survival of Listeria monocytogenes in soft cheeses

The purpose of this study was to determine the effect of sodium lactate and sodium propionate, both in combination with sodium acetate, on strains of Listeria monocytogenes in artificially inoculated soft cheeses. Minas Frescal and Coalho cheeses, inoculated with a mix of L. monocytogenes 1/2a and Scott A, underwent two treatments: 2% (w/v) sodium lactate in combination with 0.25% (w/v) sodium acetate and 2% (w/v) sodium propionate in combination with 0.25% (w/v) sodium acetate. The samples were analysed immediately and after 7 days at 10 °C. The growth of the pathogen was inhibited in cheeses containing the salts of organic acids, and the effects of treatment were statistically significant (P < 0.05). However, there was no difference between the types of treatment applied. Our data demonstrate that the effectiveness of the salts of organic acids depended on the initial concentration of L. monocytogenes and that a higher concentration of the salts is necessary to ensure sustained inactivation of target pathogens because they are weakly antilisterial when the soft cheeses are stored at 10 °C.     

Classification of Pecorino cheeses produced in Italy according to their ripening time and manufacturing technique using Fourier transform infrared spectroscopy

Fourier-transform infrared spectroscopy, followed by linear discriminant analysis of the spectral data, was used to classify Italian Pecorino cheeses according to their ripening time and manufacturing technique. The Fourier transform infrared spectra of the cheeses were divided into 18 regions and the normalized absorbance peak areas within these regions were used as predictors. Linear discriminant analysis models were constructed to classify Pecorino cheeses according to different ripening stages (hard and semi-hard) or according to their manufacturing technique (fossa and nonfossa cheeses). An excellent resolution was achieved according to both ripening time and manufacturing technique. Also, a final linear discriminant analysis model considering the 3 categories (hard nonfossa, hard fossa, and semi-hard nonfossa) was constructed. A good resolution among the 3 categories was obtained.     

The efficacy of nisin can drastically vary when produced in situ in model cheeses

Nisin, a bacteriocin produced by strains of Lactococcus lactis, has a broad inhibitory effect against Gram-positive bacteria. This study investigated the efficacy of nisin Z against Lactobacillus sakei when produced by a nisin-producing strain L. lactis in model cheeses manufactured with ultrafiltrated milk. These cheeses, containing 0, 4 or 10% of gelatin in their dry matter, were inoculated with both strains. Measurement of Lb. sakei loss of viability was an indirect indicator of nisin in situ efficacy. After 24 h, the loss of viability of Lb. sakei was from 0.73 ± 0.14 to 3.30 ± 0.60 log₁₀ cfu g⁻¹ in the cheeses with 0 and 10% of gelatin, respectively, indicating a better in situ efficacy of nisin when gelatin was incorporated. However, the concentration of nisin produced by Lactococcus was similar (3.5 μg g⁻¹) in all model cheeses when measured using an enzyme-linked immune sorbent assay (ELISA). The growth of Lactococcus was slightly improved when gelatin was incorporated, leading to a higher lactate concentration, which is one of the factors explaining the increased nisin efficacy. These results reinforced previous observations that prediction of nisin efficacy in complex food systems remains difficult.     

Prevalence,characterization and antibiotic resistance of enterococci from traditional cheeses in Turkey

The aim of the present work is to provide information about Enterococcus strains isolated from traditional Turkish cheese samples in Ankara (Turkey), focusing on their prevalence, phenotypic and genotypic characteristics, and antibiotic resistance. A total of 213 probable enterococcal isolates isolated from 215 samples were identified by phenotypic and genotypic methods. As a result of 16S rDNA sequence analysis, 88 of the 213 enterococci strains were identified as Enterococcus faecium and 125 as Enterococcus faecalis. The E. faecalis strains (58.7%) were identified as the dominant species isolated from cheese samples in Turkey. The 213 Enterococcus strains were tested for susceptibility to 12 different antimicrobial agents. The resistance phenotype were as follow: nalidixic acid (100%), kanamycin (98.6%), rifampicin (78.4%), ampicillin (48.8%), ciprofloxacin (45.5%), erythromycin (18.8%), tetracycline (11.7%), penicillin G (5.6%), chloramphenicol (4.2%), gentamycin (3.8%) and streptomycin (1.4%). None of the strains was resistant to vancomycin. E. faecium strains showed more resistant phenotypes than E. faecalis strains as shown by the antibiotic resistance levels. It was also observed that the resistance of E. faecium and E. faecalis strains against the antibiotics was statistically significant (p ? 0.05). In total, 100% of E. faecium and 88.8% of E. faecalis strains were resistant to multiple drugs.     

Shelf life and quality of probiotic yogurt produced with Lactobacillus acidophilus and Gobdin

This study evaluated the effect of dry white mulberry and walnut paste (Gobdin, a traditional Turkish food) in probiotic yogurt on the survival of Lactobacillus acidophilus and yogurt properties. Six different yogurts were produced with 0%, 5% and 10% Gobdin using Lactobacillus bulgaricus + Streptococcus thermophilus and with 0%, 5% and 10% Gobdin using L. bulgaricus + S. thermophilus + L. acidophilus. The physical, chemical, microbiological and sensorial properties of the yogurts were evaluated based on storage at 4 ± 1 °C. Probiotic shelf life and the most suitable combinations were determined. The highest L. acidophilus count (8.65 log cfu g^?1) was found in the 5% Gobdin‐supplemented yogurt on the 7th day of storage, while the lowest count (8.11 log cfu g^?1) was found in the probiotic control yogurt on the 21st day. Although the L. acidophilus counts in the probiotic yogurts declined during storage, all values found throughout the 21‐day storage period were >8 log cfu g^?1. This is above the level necessary to provide the desired therapeutic effect in probiotic products (10⁶–10⁷ cfu g^?1). The highest overall acceptability score was obtained on the first day from the yogurt with 5% Gobdin. However, all yogurt samples had general acceptability scores between 7 and 8 points from a 9‐point maximum. Thus, this study determined that a new functional yogurt can be produced using L. acidophilus with 5% Gobdin.     

Characterization of a traditional semi-hard Italian cheese produced by soaking in wine

A traditional type of semi-hard cheese which, after a ripening period in the warehouse, is soaked in wine or fermented must, was studied. In particular, the effect of the soaking phase on the characteristics of the cheese and on the profiles of the volatile compounds was investigated. The study proved that it is the alcohols and the short-chain ethyl esters which chiefly contribute to the distinctive aromatic fraction of the product. These compounds migrate from the wine into the cheese as a result of the different concentrations existing between the two systems.     

Bacterial diversity dynamics of traditional Turkish Ezine Cheese as evaluated by PCR‐DGGE and SSCP analysis

The aim of this study was to evaluate the bacterial ecosystem of milk and Ezine cheese by PCR amplification of the V3 region of the bacterial 16S rRNA gene followed by denaturing gradient gel electrophoresis (DGGE) and by monitoring the bacterial diversity dynamics using PCR single‐strand conformation polymorphism (SSCP) analysis. PCR‐DGGE analysis revealed that 17 different bands and strains belonging to the Lactococcus lactis group and Streptococcus thermophilus were predominant during manufacturing and ripening. SSCP analysis revealed that the bacterial profiles of the two cheese samples were similar.     

Optimising detection of acidification kinetics diversity in Lactococcus lactis subsp. lactis using SDS‐PAGE protein fingerprinting as screening method

Cell‐free extracts from 157 Lactococcus lactis strains isolated from Artisan cheese were screened by protein fingerprinting previous to their technological characterisation. The strains were classified according to their electrophoretic patterns into five groups. A set of strains representing the different clusters were selected to study their acidifying activity in milk. Time and rate feature points, as well as the shape of the acidification curves, resulted in six different fermentation kinetics, mostly consistent with the electrophoretic groups. Thus, selection of native strains as starter cultures based on their acidifying activity could be optimised by protein fingerprinting.     

Conjugated Linoleic Acid in Swedish dairy products with special reference to the manufacture of hard cheeses

In this study, the concentration of conjugated linoleic acid (CLA), a natural potential anticarcinogen, in different commercial Swedish dairy products, i.e. two types of pasteurized milk, five cream products, seven fermented milk products and six types of cheeses, was determined. The concentration of CLA (cis-9,trans-11 isomer) was 5.8-5.9 mg g⁻¹ fat in pasteurized milk, 4.6-6.2 in fermented milk, 6.1-6.2 in cream products, and 5.0-7.1 in cheeses. The variation in the concentration of CLA in processed products was smaller than that previously reported in raw milk. No effect of manufacture, starter culture or ripening time on the concentration of CLA in two common types of Swedish hard cheeses (Grevé and Herrgårdsost) was observed.     

Changes in physicochemical and organoleptic properties of traditional Iranian cheese Lighvan during ripening

Lighvan cheese was studied to determine the physicochemical and biochemical changes over 90 days of ripening in brine. Acidity, pH, dry matter, fat values, lipolysis level, water‐soluble nitrogen (WSN), total nitrogen (TN), ripening index (RI), trichloroacetic acid‐soluble nitrogen (TCA‐SN) and organoleptic assessments were analysed. Dry matter and fat values decreased during ripening. Lipolysis level, RI, TCA‐SN values and salt content increased continuously until the end of the ripening period, but total nitrogen decreased throughout a 90‐day storage period. The ripening stage was the main factor affecting the cheese’s sensory properties.     

Antibacterial effects of chitosan coating containing Mentha aquatica L. essence against Escherichia coli,Staphylococcus aureus and Listeria monocytogenes in Iranian white cheese

The effect of a chitosan coating and Mentha aquatica L. essence on Iranian white cheese was investigated. Results showed 100% inhibition of Escherichia coli growth using 1.5% essence after 10 days. After 15 days of incubation, the Staphylococcus aureus population was reduced by 44.2%, 70.0%, and 88.5% using 0.5, 1.0 and 1.5% essence, respectively. After 15 days, Listeria monocytogenes growth was inhibited by 63.84%, 70.12%, and 85.9% using 0.5, 1.0, and 1.5% essence, respectively. Inhibition zone diameter studies also confirmed the antibacterial effects of applied coating against all the above‐mentioned bacteria in Iranian white cheese.     

Biotechnological characterisation of exocellular proteases produced by enological Hanseniaspora isolates

Twenty‐six enological Hanseniaspora isolates were identified by ITS PCR‐RFLP and D1/D2 domain of 26S rDNA sequencing and assayed for exocellular protease production. Based on qualitative data, six isolates, belonging to H. guilliermondii, H. valbyensis and H. occidentalis species, were selected to continue the study. Analytical procedure was optimised, and protease activities were quantified and characterised on the basis of different biotechnological factors. Protease activity was quite glucose, fructose and ethanol content independent, but divalent cation affects activity; these data support that they were aspartic proteases. The effect of 2‐mercaptoethanol suggests the importance of disulphide bonds to maintain the structure of the active centre. Our results show these enzymes could be suitable for using in biotechnological processes at neutral pH, such as cheese, bread and meat industries. This is, to our knowledge, the first work showing the induction process to induce and characterise proteolytic activity in Hanseniaspora isolates.     

Formation and dispersal of biofilms in dairy substrates

The formation and dispersion of biofilms in the dairy industry is a problem because it increases cross‐contamination, affecting the shelf life of the products and their safety. The objective of this study was to evaluate the influence of different dairy substrates (cows’ milk and whey protein) on the formation and dispersion of Enterococcus faecalis, Listeria monocytogenes, Staphylococcus aureus and Bacillus cereus in two biofilm systems (mono‐species and multi‐species) on stainless steel at 25 °C. The dominant behaviour of E. faecalis occurred in most of the tests on mono‐species and multi‐species biofilms. A greater dispersion of biofilm cells was observed in skimmed milk.