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Nisin is a bacteriocin that is globally employed as a biopreservative in food systems to control gram-positive, and some gram-negative, bacteria. Here we tested the bioactivity of nisin A-producing Lactococcus lactis NZ9700 and producers of bioengineered variants thereof against representatives of the gram-negative genus Thermus, which has been associated with the pink discoloration defect in cheese. Starting with a total of 73 nisin variant-producing Lactococcus lactis, bioactivity against Thermus was assessed via agar diffusion assays, and 22 variants were found to have bioactivity greater than or equal to that of the nisin A-producing control. To determine to what extent this enhanced bioactivity was attributable to an increase in specific activity, minimum inhibitory concentrations were determined using the corresponding purified form of these 22 nisin A derivatives. From these experiments, nisin M17Q and M21F were identified as peptides with enhanced antimicrobial activity against the majority of Thermus target strains tested. In addition, several other peptide variants were found to exhibit enhanced specific activity against a subset of strains.

peer-reviewed The objective of this study was to investigate the effect of adding different levels of a thermoresistant protease produced by a Pseudomonas fluorescens strain to milk on the manufacture and quality of Cheddar cheese. Fresh raw milk was collected, standardized, and pasteurized at 72°C for 15 s, and the enzyme was added to give a protease activity of 0.15 or 0.60 U/L (treatments P1 and P4, respectively), while one sample had no enzyme added (control). Milk was stored at 4°C for 48 h and Cheddar cheese was manufactured after 0 and 48 h of storage. Results indicated that the protease was active in milk during 48 h of storage; however, its effect on milk composition was minimal. The protein that was preferentially hydrolyzed by the protease over storage was β-casein, followed by κ-casein. The mean cheese yield and recovery of fat and protein obtained for all cheeses were not affected by protease activity. The protease showed low activity during cheese manufacture, possibly because of unfavorable conditions, including low pH. One of the factors that might have influenced protease activity was the pH of the curd (approximately 6.55 after acidification and 5.35 at milling), which was lower than that at which the enzyme would have optimum activity (pH 7 to 9). Consequently, the composition, pH, patterns of proteolysis, and hardness of all cheeses produced were similar and in accordance with values expected for that type of cheese, independently of the protease activity level. However, slight increases in proteolysis were observed in P4 cheeses and produced using milk stored for 48 h. Both the P1 and P4 cheeses had higher concentrations of free amino acids (FAA) compared with the control, whereas urea-PAGE electrophoretograms indicated a greater breakdown of caseins in the P4 cheese samples, which may be related to possible increases in numbers of proteolytic bacteria in milk during storage. Therefore, the thermoresistant psychrotrophic bacterial protease(s) tested in this study may affect the manufacture or quality of Cheddar cheese during ripening to a relatively limited extent. However, controlling initial levels of proteolytic bacteria in raw milk remains essential, because proteolysis affects the development of flavor and texture in cheese.

Carotenoids are isoprenoids widely distributed in foods that have been always part of the diet of humans. Unlike the other so-called food bioactives, some carotenoids can be converted into retinoids exhibiting vitamin A activity, which is essential for humans. Furthermore, they are much more versatile as they are relevant in foods not only as sources of vitamin A, but also as natural pigments, antioxidants, and health-promoting compounds. Lately, they are also attracting interest in the context of nutricosmetics, as they have been shown to provide cosmetic benefits when ingested in appropriate amounts. In this work, resulting from the collaborative work of participants of the COST Action European network to advance carotenoid research and applications in agro-food and health (EUROCAROTEN, www.eurocaroten.eu, https://www.cost.eu/actions/CA15136/#tabs|Name:overview) research on carotenoids in foods and feeds is thoroughly reviewed covering aspects such as analysis, carotenoid food sources, carotenoid databases, effect of processing and storage conditions, new trends in carotenoid extraction, daily intakes, use as human, and feed additives are addressed. Furthermore, classical and recent patents regarding the obtaining and formulation of carotenoids for several purposes are pinpointed and briefly discussed. Lastly, emerging research lines as well as research needs are highlighted., This article is based upon work from COST Action (European network to advance carotenoid research and applications in agro-food and health, EUROCAROTEN, CA15136, www.eurocaroten.eu, https://www.cost.eu/actions/CA15136/#tabs|Name:overview) supported by COST (European Cooperation in Science and Technology, http://www.cost.eu/).

peer-reviewed The effects of the independent variables protein concentration (4–6%), coagulum cut size (6–18 mm3), and coagulation temperature (28–36°C) on curd moisture loss during in-vat stirring were investigated using response surface methodology. Milk (14 kg) in a cheese vat was rennet coagulated, cut, and stirred as per semihard cheesemaking conditions. During stirring, the moisture content of curd samples was determined every 10 min between 5 and 115 min after cutting. The moisture loss kinetics of curds cut to 6 mm3 followed a logarithmic trend, but the moisture loss of curds from larger cut sizes, 12 or 18 mm3, showed a linear trend. Response surface modeling showed that curd moisture level was positively correlated with cut size and negatively correlated with milk protein level. However, coagulation temperature had a significant negative effect on curd moisture up to 45 min of stirring but not after 55 min (i.e., after cooking). It was shown that curds set at the lower temperature had a slower syneresis rate during the initial stirring compared with curds set at a higher temperature, which could be accelerated by reducing the cut size. This study shows that keeping a fixed cut size at increasing protein concentration decreased the level of curd moisture at a given time during stirring. Therefore, to obtain a uniform curd moisture content at a given stirring time at increasing protein levels, an increased coagulum cut size is required. It was also clear that breakage of the larger curd particles during initial stirring can also significantly influence the curd moisture loss kinetics. Both transmission and scanning electron micrographs of cooked curds (i.e., after 45 min of stirring) showed that the casein micelles were fused at a higher degree in curds coagulated at 36°C compared with 28°C, which confirmed that coagulation temperature causes a marked change in curd microstructure during the earlier stages of stirring. The present study showed the dynamics of curd moisture content during stirring when using protein-concentrated milk at various set temperatures and cut sizes. This provides the basis for achieving a desired curd moisture loss during cheese manufacture using protein-concentrated milk as a means of reducing the effect of seasonal variation in milk for cheesemaking.

This study investigated the effect of centrifugation (9,000 × g, 50°C, flow rate = 1,000 L/h), as well as the incorporation of high-heat-treated (HHT) centrifugate into cheese milk on the composition, texture, and ripening characteristics of Maasdam cheese. Neither centrifugation nor incorporation of HHT centrifugate into cheese milk had a pronounced effect on the compositional parameters of any experimental cheeses, except for moisture and moisture in nonfat substance (MNFS) levels. Incorporation of HHT centrifugate at a rate of 6 to 10% of the total milk weight into centrifuged milk increased the level of denatured whey protein in the cheese milk and also increased the level of MNFS in the resultant cheese compared with cheeses made from centrifuged milk and control cheeses; moreover, cheese made from centrifuged milk had ∼3% higher moisture content on average than control cheeses. Centrifugation of cheese milk reduced the somatic cell count by ∼95% relative to the somatic cell count in raw milk. Neither centrifugation nor incorporation of HHT centrifugate into cheese milk had a significant effect on age-related changes in pH, lactate content, and levels of primary and secondary proteolysis. However, the value for hardness was significantly lower for cheeses made from milk containing HHT centrifugate than for other experimental cheese types. Overall, centrifugation appeared to have little effect on composition, texture, and ripening characteristics of Maasdam cheese. However, care should be taken when incorporating HHT centrifugate into cheese milk, because such practices can influence the level of moisture, MNFS, and texture (particularly hardness) of resultant cheeses. Such differences may have the potential to influence subsequent eye development characteristic, although no definitive trends were observed in the present study and further research on this is recommended.

The effect of varying cook temperature (40 or 50 °C) and salting method (dry or brine salting) on bacterial viability, enzymatic activity and chemical composition in cheeses made using Streptococcus thermophilus and Lactobacillus helveticus were investigated. Dry salting resulted in decreased cell viability of L. helveticus, increased lactate dehydrogenase activity and increased free amino acid levels, compared with brine salted cheeses, irrespective of cook temperature. A cook temperature of 50 °C resulted in reduced primary proteolysis and increased pH in comparison with that in cheeses cooked to 40 °C. Salting method influenced moisture content with higher levels in brine salted cheeses; cook temperature was also influential with higher cook temperature resulting in lower moisture within each salting method. Variations in manufacture procedure can allow for the development of cheese varieties with novel flavour and texture profiles using existing Cheddar or Swiss-style manufacturing facilities.

The individual roles of hydrolysis of αS1- and β-caseins, and calcium solubilization on the fracture properties of semi-hard cheeses, such as Maasdam and other eye-type cheeses, remain unclear. In this study, the hydrolysis patterns of casein were selectively altered by adding a chymosin inhibitor to the curd/whey mixture during cheese manufacture, by substituting fermentation-produced bovine chymosin (FPBC) with fermentation-produced camel chymosin (FPCC), or by modulating ripening temperature. Moreover, the level of insoluble calcium during ripening was quantified in all cheeses. Addition of a chymosin inhibitor, substitution of FPBC with FPCC, or ripening of cheeses at a consistent low temperature (8 °C) decreased the hydrolysis of αS1-casein by ~95%, ~45%, or ~30%, respectively, after 90 d of ripening, whereas ~35% of β-casein was hydrolysed in that time for all cheeses, except for those ripened at a lower temperature (~17%). The proportion of insoluble calcium as a percentage of total calcium decreased significantly from ~75% to ~60% between 1 and 90 d. The rigidity or strength of the cheese matrix was found to be higher (as indicated by higher fracture stress) in cheeses with lower levels of proteolysis or higher levels of intact caseins, primarily αS1-casein. However, contrary to the expectation that shortness of cheese texture is associated with αS1-casein hydrolysis, fracture strain was significantly positively correlated with the level of intact β-casein and insoluble calcium content, indicating that the cheeses with low levels of intact β-casein or insoluble calcium content were more likely to be shorter in texture (i.e., lower fracture strain). Overall, this study suggests that the fracture properties of cheese can be modified by selective hydrolysis of caseins, altering the level of insoluble calcium or both. Such approaches could be applied to design cheese with specific properties.

The objective of this study was to identify the carotenoids imparting the orange colour to the rind, and pale yellow color to the core, of selected smear-ripened cheeses. The cheeses investigated were Charloe, Ashbrook, Taleggio, and Limburger, and were sourced from artisanal markets. Samples of the rind and core were extracted using non-polar solvents, followed by saponification to hydrolyze triglycerides to remove fatty acids, and to release carotenoid esters. Extracts were tested using ultra-high pressure liquid chromatograph-diode array detector-high resolution mass spectrometry (UHPLC-DAD-MS and -MS/MS), and identities of α- and β-carotene, lycopene, and β-cryptoxanthin confirmed with authentic standards. β-Carotene was the predominant species in both the rind and core, absorbing ~70% of the signal at 450 nm in all cheese extracts tested, as well as minor quantities of β-cryptoxanthin and α-carotene. Carotenoids unique to the rind included lycopene as well as the rare bacterial carotenoids previously identified in bacterial isolates of cheeses (i.e. decaprenoxanthin, sarcinaxanthin, and echinenone). This is the first detailed characterisation of carotenoids extracted directly from smear-ripened cheeses, and reveals that smear-ripened cheese can contribute both provitamin A carotenoids as well as C50 carotenoids to the human diet.

Nonstarter lactic acid bacteria are commonly implicated in undesirable gas formation in several varieties, including Cheddar, Dutch-, and Swiss-type cheeses, primarily due to their ability to ferment a wide variety of substrates. This effect can be magnified due to factors that detrimentally affect the composition or activity of starter bacteria, resulting in the presence of greater than normal amounts of fermentable carbohydrates and citrate. The objective of this study was to determine the potential for a facultatively heterofermentative Lactobacillus (Lactobacillus casei DPC6987) isolated from a cheese plant environment to promote gas defects in the event of compromised starter activity. A Swiss-type cheese was manufactured, at pilot scale and in triplicate, containing a typical starter culture (Streptococcus thermophilus and Lactobacillus helveticus) together with propionic acid bacteria. Lactobacillus helveticus populations were omitted in certain vats to mimic starter failure. Lactobacillus casei DPC6987 was added to each experimental vat at 4 log cfu/g. Cheese compositional analysis and X-ray computed tomography revealed that the failure of starter bacteria, in this case L. helveticus, coupled with the presence of a faculatively heterofermentative Lactobacillus (L. casei) led to excessive eye formation during ripening. The availability of excess amounts of lactose, galactose, and citrate during the initial ripening stages likely provided the heterofermentative L. casei with sufficient substrates for gas formation. The accrual of these fermentable substrates was notable in cheeses lacking the L. helveticus starter population. The results of this study are commercially relevant, as they demonstrate the importance of viability of starter populations and the control of specific nonstarter lactic acid bacteria to ensure appropriate eye formation in Swiss-type cheese.

A cascade membrane filtration process including microfiltration (MF), ultrafiltration (UF) and reverse osmosis (RO) was used to fractionate skim milk into different streams. Significant quantities of lactose and minerals were removed to permeate after MF at 0.14 μm. Cheese milk, of similar casein content to the raw milk, was standardised simultaneously for casein, lactose, ash and total calcium from the membrane streams without requiring CaCl2 and lactose addition. Serum protein depleted cheese milk of typical casein content had similar rennet coagulability, cheese composition, texture and yield to the control; milk of 1.5 × casein content had a faster coagulation rate and resulted in cheese of lower moisture content. On a dry matter basis, the serum protein content of MF permeate concentrated by UF was significantly higher than that in cheese whey (51.54% versus 5.63–9.45%), with significantly lower contents of ash (0.95% versus 7.11–7.53%) and lactose (9.50% versus 61.98–70.35%).

peer-reviewed This study characterized the coagulation properties and defined the cutting window (CW; time between storage modulus values of 35 and 70 Pa) using rheometry for milk standardized to 4, 5, or 6% protein and set at 28, 32, or 36°C. Milks were standardized to a protein-to-fat ratio of approximately 1 by blending ultrafiltration retentate, skim milk, and whole milk. The internal curd microstructure for selected curd samples was analyzed with transmission electron microscopy and scanning electron microscopy. Lowering the coagulation temperature caused longer rennet coagulation time and time to reach storage modulus of 35 Pa, translating into a wider CW. It also led to a lower maximum curd-firming rate (MCFR) with lower firmness at 40 min at a given protein level. Increasing protein levels resulted in the opposite effect, although without an effect on rennet coagulation time at a given temperature. On coagulation at 28°C, milk with 5% protein resulted in a similar MCFR (∼4 Pa/min) and CW (∼8.25 min) compared with milk with 4% protein at 32°C, which reflects more standard conditions, whereas increasing milk to 6% protein resulted in more than doubling of the curd-firming rate (MCFR = 9.20 Pa/min) and a shorter CW (4.60 min). Gels set at 28°C had lower levels of rearrangement of protein network after 40 min compared with those set at 36°C. Protein levels, on the other hand, had no influence on the levels of protein network rearrangement, as indicated by loss tangent values. The internal structure of curd particles, as investigated by both scanning electron microscopy and transmission electron microscopy, appeared to have less cross-linking and smaller casein aggregates when coagulated at 28°C compared with 36°C, whereas varying protein levels did not show a marked effect on aggregate formation. Overall, this study showed a marked interactive effect between coagulation temperature and protein standardization of milk on coagulation properties, which subsequently requires adjustment of the CW during cheesemaking. Lowering of the coagulation temperature greatly altered the curd microstructure, with a tendency for less syneresis during cutting. Further research is required to quantify the changes in syneresis and in fat and protein losses to whey due to changes in the microstructure of curd particles arising from the different coagulation conditions applied to the protein-fortified milk.

peer-reviewed The non-starter microbiota of Cheddar cheese mostly comprises mesophilic lactobacilli, such as Lactobacillus casei, Lactobacillus paracasei, Lactobacillus rhamnosus, and Lactobacillus plantarum. These bacteria are recognized for their potential to improve Cheddar cheese flavor when used as adjunct cultures. In this study, three strains of L. paracasei (DPC2071, DPC4206, and DPC4536) were evaluated for their contribution to the enhancement and diversification of flavor in short-aged Cheddar cheese. The strains were selected based on their previously determined genomic diversity, variability in proteolytic enzyme activities and metabolic capability in cheese model systems. The addition of adjunct cultures did not affect the gross composition or levels of lipolysis of the cheeses. The levels of free amino acids (FAA) in cheeses showed a significant increase after 28 days of ripening. However, the concentrations of individual amino acids in the cheeses did not significantly differ except for some amino acids (aspartic acid, threonine, serine, and tryptophan) at Day 14. Volatile profile analysis revealed that the main compounds that differentiated the cheeses were of lipid origin, such as long chain aldehydes, acids, ketones, and lactones. This study demonstrated that the adjunct L. paracasei strains contributed to the development and diversification of compounds related to flavor in short-aged Cheddar cheeses.

peer-reviewed The microbiota of cheese plays a key role in determining its organoleptic and other physico-chemical properties. It is essential to understand the various contributions, positive or negative, of these microbial components in order to promote the growth of desirable taxa and, thus, characteristics. The recent application of high throughput DNA sequencing (HTS) facilitates an even more accurate identification of these microbes, and their functional properties, and has the potential to reveal those microbes, and associated pathways, responsible for favorable or unfavorable characteristics. This technology also facilitates a detailed analysis of the composition and functional potential of the microbiota of milk, curd, whey, mixed starters, processing environments, and how these contribute to the final cheese microbiota, and associated characteristics. Ultimately, this information can be harnessed by producers to optimize the quality, safety, and commercial value of their products. In this review we highlight a number of key studies in which HTS was employed to study the cheese microbiota, and pay particular attention to those of greatest relevance to industry.

The non-starter microbiota of Cheddar cheese mostly comprises mesophilic lactobacilli, such as Lactobacillus casei, Lactobacillus paracasei, Lactobacillus rhamnosus, and Lactobacillus plantarum. These bacteria are recognized for their potential to improve Cheddar cheese flavor when used as adjunct cultures. In this study, three strains of L. paracasei (DPC2071, DPC4206, and DPC4536) were evaluated for their contribution to the enhancement and diversification of flavor in short-aged Cheddar cheese. The strains were selected based on their previously determined genomic diversity, variability in proteolytic enzyme activities and metabolic capability in cheese model systems. The addition of adjunct cultures did not affect the gross composition or levels of lipolysis of the cheeses. The levels of free amino acids (FAA) in cheeses showed a significant increase after 28 days of ripening. However, the concentrations of individual amino acids in the cheeses did not significantly differ except for some amino acids (aspartic acid, threonine, serine, and tryptophan) at Day 14. Volatile profile analysis revealed that the main compounds that differentiated the cheeses were of lipid origin, such as long chain aldehydes, acids, ketones, and lactones. This study demonstrated that the adjunct L. paracasei strains contributed to the development and diversification of compounds related to flavor in short-aged Cheddar cheeses.

Fermented foods, in particular, surface-ripened cheese, represent a model to explain the metabolic interactions which regulate microbial succession in complex environments. This study explains the role of individual species in a heterogeneous microbial environment, i.e., the exterior of surface-ripened cheese. Through whole-metagenome shotgun sequencing, it was possible to investigate the metabolic potential of the resident microorganisms and show how variations in the microbial populations influence important aspects of cheese ripening, especially flavor development. Overall, in addition to providing fundamental insights, this research has considerable industrial relevance relating to the production of fermented food with specific qualities., In this study, a young Cheddar curd was used to produce two types of surface-ripened cheese, using two commercial smear-culture mixes of yeasts and bacteria. Whole-metagenome shotgun sequencing was used to screen the microbial population within the smear-culture mixes and on the cheese surface, with comparisons of microorganisms at both the species and the strain level. The use of two smear mixes resulted in the development of distinct microbiotas on the surfaces of the two test cheeses. In one case, most of the species inoculated on the cheese established themselves successfully on the surface during ripening, while in the other, some of the species inoculated were not detected during ripening and the most dominant bacterial species, Glutamicibacter arilaitensis, was not a constituent of the culture mix. Generally, yeast species, such as Debaryomyces hansenii and Geotrichum candidum, were dominant during the first stage of ripening but were overtaken by bacterial species, such as Brevibacterium linens and G. arilaitensis, in the later stages. Using correlation analysis, it was possible to associate individual microorganisms with volatile compounds detected by gas chromatography-mass spectrometry in the cheese surface. Specifically, D. hansenii correlated with the production of alcohols and carboxylic acids, G. arilaitensis with alcohols, carboxylic acids and ketones, and B. linens and G. candidum with sulfur compounds. In addition, metagenomic sequencing was used to analyze the metabolic potential of the microbial populations on the surfaces of the test cheeses, revealing a high relative abundance of metagenomic clusters associated with the modification of color, variation of pH, and flavor development. IMPORTANCE Fermented foods, in particular, surface-ripened cheese, represent a model to explain the metabolic interactions which regulate microbial succession in complex environments. This study explains the role of individual species in a heterogeneous microbial environment, i.e., the exterior of surface-ripened cheese. Through whole-metagenome shotgun sequencing, it was possible to investigate the metabolic potential of the resident microorganisms and show how variations in the microbial populations influence important aspects of cheese ripening, especially flavor development. Overall, in addition to providing fundamental insights, this research has considerable industrial relevance relating to the production of fermented food with specific qualities.

peer-reviewed Centrifugation is a common milk pretreatment method for removal of Clostridium spores which, on germination, can produce high levels of butyric acid and gas, resulting in rancid, gassy cheese. The aim of this study was to determine the effect of centrifugation of milk, as well as incorporation of high heat-treated centrifugate into cheese milk, on the microbial and volatile profile of Maasdam cheese. To facilitate this, 16S rRNA amplicon sequencing in combination with a selective media-based approach were used to study the microbial composition of cheese during maturation, and volatile organic compounds within the cheese matrix were analyzed by HPLC and solid-phase microextraction coupled with gas chromatography–mass spectrometry. Both culture-based and molecular approaches revealed major differences in microbial populations within the cheese matrix before and after warm room ripening. During warm room ripening, an increase in counts of propionic acid bacteria (by ∼101.5 cfu) and nonstarter lactic acid bacteria (by ∼108 cfu) and a decrease in the counts of Lactobacillus helveticus (by ∼102.5 cfu) were observed. Lactococcus species dominated the curd population throughout ripening, followed by Lactobacillus, Propionibacterium, and Leuconostoc, and the relative abundance of these accounted for more than 99% of the total genera, as revealed by high-throughput sequencing. Among subdominant microflora, the overall relative abundance of Clostridium sensu stricto was lower in cheeses made from centrifuged milk than control cheeses, which coincided with lower levels of butyric acid. Centrifugation as well as incorporation of high heat-treated centrifugate into cheese milk seemed to have little effect on the volatile profile of Maasdam cheese, except for butyric acid levels. Overall, this study suggests that centrifugation of milk before cheesemaking is a suitable method for controlling undesirable butyric acid fermentation without significantly altering the levels of other volatile organic compounds of Maasdam cheese.

peer-reviewed The influence of buttermilk or buttermilk powder addition to cheese milk or cheese curds respectively on cheese functional properties, free fatty acid profiles and subsequent volatile and sensory characteristics was investigated. Buttermilk addition to cheese milk resulted in a softer cheese compared to other cheeses, with a significantly reduced flowability, while buttermilk powder addition had no influence on cheese firmness but cheese flowability was also reduced compared to the control cheese. Larger pools of free fat, higher levels of free fatty acids, volatile compounds and significant differences in sensory profiles associated with off-flavour were also observed with the addition of buttermilk to cheese milk. Application of light microscopy, using toluidine blue stain, facilitated the visualisation of fat globule structure and distribution within the protein matrix. Addition of 10% buttermilk powder resulted in significant increases in volatile compounds originating from proteolysis pathways associated with roasted, green aromas. Descriptive sensory evaluation indicated few differences between the 10% buttermilk powder and the control cheese, while buttermilk cheeses scored negatively for sweaty, barnyard aromas, oxidized and off flavors, correlating with associated volatile aromas. Addition of 10% buttermilk powder to cheese curds results in cheese comparable to the control Cheddar with some variations in volatile compounds resulting in a cheese with similar structural and sensory characteristics albeit with subtle differences in overall cheese flavor. This could be manipulated to produce cheeses of desirable quality, with potential health benefits due to increased phospholipid levels in cheese.

The strains Staphylococcus saprophyticus DPC5671 and Corynebacterium casei DPC5298 were applied in combination with Debaryomyces hansenii DPC6258 to the surface of young Cheddar cheese curd to obtain two different smear-ripened cheeses. A surface microbiota developed over the incubation period, comprising of both yeast and bacteria; pulsed field gel electrophoresis confirmed that the inoculated strains of S. saprophyticus DPC5671 or C. casei DPC5298 were the dominant bacterial strains on the surface of the cheese at the end of the ripening period. The smear cultures changed the appearance and aroma, which were significantly different from the control cheese. The approach presented in this study represents a method for the development of new cheese varieties with novel aromas within a short ripening time.

The quality of any type of cheese is without doubt influenced by the quality of the milk from which it is made, from microbiological, biochemical, sensory, and many other perspectives. Raw milk is an inherently variable starting point for cheese manufacture, with its composition, for example, being influenced by factors, such as stage of lactation, diet, and health status of the producing animal, which subsequently determines milk coagulation properties and cheese yield, composition and likely ripening. Protein standardization treatments may be applied to cheese milks to minimize resultant cheese compositional variability. Similarly, milk is also biochemically complex, with a wide range of indigenous enzymes, some of which, for example, plasmin and lipase, are variable in activity and susceptible to processes applied to the milk, while contributing to cheese ripening. Finally, raw milk may act as a vector for bacterial spores and also provides an excellent medium for growth of a wide range of microorganisms. As cheese milk is vulnerable to contamination at multiple points before entering the cheese vat, control of the same for cheese safety and quality is critical.

The purpose of this study was to investigate the effects of pasture-based versus indoor total mixed ration (TMR) feeding systems on the chemical composition, quality characteristics, and sensory properties of full-fat Cheddar cheeses. Fifty-four multiparous and primiparous Friesian cows were divided into 3 groups (n = 18) for an entire lactation. Group 1 was housed indoors and fed a TMR diet of grass silage, maize silage, and concentrates; group 2 was maintained outdoors on perennial ryegrass only pasture (GRS); and group 3 was maintained outdoors on perennial ryegrass/white clover pasture (CLV). Full-fat Cheddar cheeses were manufactured in triplicate at pilot scale from each feeding system in September 2015 and were examined over a 270-d ripening period at 8°C. Pasture-derived feeding systems were shown to produce Cheddar cheeses yellower in color than that of TMR, which was positively correlated with increased cheese β-carotene content. Feeding system had a significant effect on the fatty acid composition of the cheeses. The nutritional composition of Cheddar cheese was improved through pasture-based feeding systems, with significantly lower thrombogenicity index scores and a greater than 2-fold increase in the concentration of vaccenic acid and the bioactive conjugated linoleic acid C18:2 cis-9,trans-11, whereas TMR-derived cheeses had significantly higher palmitic acid content. Fatty acid profiling of cheeses coupled with multivariate analysis showed clear separation of Cheddar cheeses derived from pasture-based diets (GRS or CLV) from that of a TMR system. Such alterations in the fatty acid profile resulted in pasture-derived cheeses having reduced hardness scores at room temperature. Feeding system and ripening time had a significant effect on the volatile profile of the Cheddar cheeses. Pasture-derived Cheddar cheeses had significantly higher concentrations of the hydrocarbon toluene, whereas TMR-derived cheese had significantly higher concentration of 2,3-butanediol. Ripening period resulted in significant alterations to cheese volatile profiles, with increases in acid-, alcohol-, aldehyde-, ester-, and terpene-based volatile compounds. This study has demonstrated the benefits of pasture-derived feeding systems for production of Cheddar cheeses with enhanced nutritional and rheological quality compared with a TMR feeding system.

Split and secondary fermentation defects in Swiss-type cheese varieties are manifested as undesirable slits or cracks that may lead to downgrading of the cheese. Split defect is associated with an excessive production of gas or an unsuitable cheese body that cannot accommodate gas produced, or a combination of both factors. Secondary fermentation is the apparent production of gas after the desired propionic fermentation of the warm room has taken place. No consensus exists as to the definitive causes of the defects, but possible causes are reviewed under factors that are associated with rheological behaviour (including cheese manufacture, acidification, intact protein content and proteolysis, seasonality of milk supply and ripening or storage temperature and duration) or with overproduction of gas (including milk microflora, propionic acid bacteria (PAB) — in particular strains with high aspartase activity and ability to grow at low temperatures, lactic acid bacteria, interactions between starter bacteria, facultatively heterofermentative lactobacilli (FHL) and other sources of gas including butyric acid fermentation). The influence of other parameters such as copper concentration, air incorporation, salt content, rind formation and cheese wrapping materials is also considered. Methods to reduce the prevalence of the split defect and secondary fermentation include addition of water to improve elastic properties by the removal of unfermented carbohydrate and the use of FHL to control PAB activity to prevent the production of excessive gas.

Semi-hard cheeses were manufactured from caprine milk and bovine milk mixed in ratios of 100:0; 75:25; 50:50; 25:75; 0:100 (caprine:bovine) and ripened at 12 °C for 150 d. Substitution of up to 75% bovine for caprine milks had no significant effect on levels of cheese protein, moisture-in-non-fat substances, salt, pH, whiteness, total or individual free amino acids and substitution of up to 50% had no significant effect on levels of primary proteolysis and free fatty acid and fruity flavours. Increased bovine milk content resulted in decreased levels of butanoic to octanoic acids, ethyl and methyl esters of acetic to octanoic acids and acyclic, mono and bicyclic terpenes and waxy/goaty flavours to a degree dependent on the proportion of each milk type. Partial substitution of caprine with bovine milks resulted in cheeses with chemical and sensorial characteristics that may provide an available alternative to consumers desiring caprine cheeses.

The influence of varying process (curd washing, drain pH, dry or brine salting) and ripening (at 9 or 12°C without a hot room step) parameters on manufacture and on biochemical changes during ripening of semi-hard cheeses with propionic acid bacteria (PAB) was studied. Levels of secondary proteolysis were significantly higher and viable thermophilic culture counts decreased at a faster rate in the dry-salted cheeses compared with the brine-salted cheeses. Increased ripening time and temperature resulted in increased PAB counts, levels of acetate, propionate, primary and secondary proteolysis. Increased cheese pH was significantly correlated with increased PAB counts during ripening. The washed-curd, dry-salted cheeses at 182 d of ripening at 12°C resulted in counts of PAB and levels of acetate and propionate similar to those of Swiss-type control cheeses ripened with a hot room temperature. The approach described may be used to manufacture Swiss-type cheese using technologies associated with Cheddar-type manufacture.

Semi-hard cheeses were manufactured using Streptococcus thermophilus and Lactobacillus helveticus cultures and their ripening was characterised. During cheese manufacture, curds were cooked to a maximum temperature of 47, 50 or 53 °C, pre-pressed under whey at pH 6.15, moulded, pressed and brined. Increased cook temperature resulted in increased manufacture time, a significantly reduced growth rate of S. thermophilus during manufacture in the order 47≈50 °C>53 °C and in significantly lower mean viable cell counts of S. thermophilus up to 56 d of ripening. Increasing cook temperature had no significant effect on mean viable cell numbers of L. helveticus or non-starter lactic acid bacteria (NSLAB). Cheeses produced from curds cooked to 47 °C had significantly higher levels of moisture in non-fat substances (MNFSs), salt-in-moisture and a significantly lower pH and levels of butyrate compared with cheeses produced from curds cooked to 50 or 53 °C.

Exopolysaccharide-producing Lactobacillus mucosae DPC 6426 was previously shown to have promising hypocholesterolemic activity in the atherosclerosis-prone apolipoprotein-E-deficient (apoE(-/-)) murine model. The aim of this study was to investigate the suitability of reduced-fat Cheddar and Swiss-type cheeses as functional (carrier) foods for delivery of this probiotic strain. All cheeses were manufactured at pilot-scale (500-L vats) in triplicate, with standard commercially available starters: for Cheddar, Lactococcus lactis; and for Swiss-type cheese, Streptococcus thermophilus, Lactobacillus helveticus, and Propionibacterium freudenreichii. Lactobacillus mucosae DPC 6426 was used as an adjunct culture during cheese manufacture, at a level of ~10(6) cfu·mL(-1) cheese milk (subsequently present in the cheese curd at>10(7) cfu·g(-1)). The adjunct strain remained viable at >5×10(7) cfu·g(-1) in both Swiss-type and Cheddar cheeses following ripening for 6 mo. Sensory analysis revealed that the presence of the adjunct culture imparted a more appealing appearance in Swiss-type cheese, but had no significant effect on the sensory characteristics of Cheddar cheeses. Moreover, the adjunct culture had no significant effect on cheese composition, proteolysis, pH, or instrumentally quantified textural characteristics of Cheddar cheeses. These data indicate that low-fat Swiss-type and Cheddar cheeses represent suitable food matrices for the delivery of the hypocholesterolemic Lactobacillus mucosae DPC 6426 in an industrial setting.

peer-reviewed The growth, location, and distribution of bacterial colonies in dairy products are important factors for the ripening and flavor development of cheeses, yogurts, and soured creams. Starter, non-starter, spoilage, and pathogenic bacteria all become entrapped in the developing casein matrix of dairy foods. In order to visualize these bacterial colonies and the environments surrounding them, microscopy techniques are used. The use of various microscopy methods allow for the rapid detection, enumeration, and distribution of starter, non-starter and pathogenic bacteria in dairy foods. Confocal laser scanning microscopy is extensively utilized to identify bacteria location via the use of fluorescent dyes. Further study is needed in relation to the development of micro- gradients and localized ripening parameters in dairy products due to the location of bacteria at the protein-fat interface. Development in the area of bacterial discrimination using microscopy techniques and fluorescent dyes/tags is needed as the benefits of rapidly identifying spoilage/pathogenic bacteria early in product manufacture would be of huge benefit in relation to both safety and financial concerns.

Smear-ripened cheese varieties are characterised by the growth of a smear culture, con- taining predominantly Brevibacterium linens, on the cheese surface during ripening. In such cheese, considerable zonal differences in biochemistry of ripening exist, due to moisture loss from, and growth and metabolic activity of smear microflora at, the cheese surface. In this study, the effects of adding exogenous plasmin or small amounts of mastitic milk to good quality milk on the quality of smear-ripened cheese made subsequently was examined. Addition of plasmin did not influence cheese composition immediately after manufacture, but slightly decreased the rate of moisture loss during cheese ripening. Plasmin activity decreased during the early stages of ripening, but subse- quently increased towards the end of ripening, perhaps due to changing pH conditions in the cheese. Addition of plasmin increased rates of primary proteolysis in cheese, as measured by levels of pH 4.6- soluble N and urea-PAGE, although production of later products of proteolysis appeared less af- fected. Addition of mastitic milk had largely similar effects to addition of exogenous plasmin, which may reflect a high content of plasmin or plasminogen activators in such milk. Overall, changes in milk quality and enzymology appear to influence the quality of smear-ripened cheese. Plasmin / cheese / smear-ripened

Classically, very few pre-treatments are applied to milk for cheese-making, with some cheese varieties simply made from raw whole milk, but most made from pasteurised milk of which the composition (e.g., fat:protein ratio) may have been standardized. However, there has been consistent interest in more novel and sophisticated strategies for pre-treatment of cheese-milk. Approaches explored include the use of alternative processing technologies (e.g., membrane filtration, high-pressure treatment, homogenisation, heat treatments more severe than pasteurisation) or addition of sources of protein or milk solids (e.g., milk powders, whey protein products) or enzymes. The principal reasons for such pre-treatments of cheese-milk are: (1) to control the microbiology of the raw milk and the resulting cheese better than is possible by pasteurisation (e.g., inactivation or removal of spores, control of non-starter lactic acid bacteria); (2) increasing the yield of cheese, e.g., through heat- or pressure-induced incorporation of whey proteins, or enhancing sensory properties of reduced fat cheese by direct addition of microparticulated whey proteins; (3) manipulation of cheese ripening, e.g., reducing the likelihood of off-flavour development by inactivation of enzymes or accelerating ripening through increasing enzyme-substrate interactions; or (4) improving the texture and other functional properties, e.g., melting. Finally, the considerations for manufacture and ripening of different cheese varieties, or sub-classes of specific varieties (e.g., low-fat cheese) will clearly differ and add to the complexity of the technological options available. This article will review the key principles for pre-treatment of cheese-milk, as summarised briefly above.