Queso Chihuahua: effects of seasonality of cheesemilk on rheology†

Fresh semihard raw milk (RM) and pasteurized milk (PM) cheeses made by Mennonite communities in Chihuahua, Mexico were sampled in early winter, mid‐spring and late summer and evaluated during storage to determine if rheological properties were affected by the season the cheese was produced. Seasonal effects were observed in the rheology of the RM cheeses, which underwent more extensive proteolysis than the PM cheeses. The rheological properties of the PM cheeses and the composition of all cheeses were consistent throughout the year. The texture of the RM cheese is affected by season, whereas the variability in texture and composition of PM cheese is minimal, allowing for the manufacture of a uniform product.     

Nonstarter lactic acid bacteria biofilms and calcium lactate crystals in Cheddar cheese

A sanitized cheese plant was swabbed for the presence of nonstarter lactic acid bacteria (NSLAB) biofilms. Swabs were analyzed to determine the sources and microorganisms responsible for contamination. In pilot plant experiments, cheese vats filled with standard cheese milk (lactose:protein = 1.47) and ultrafiltered cheese milk (lactose:protein = 1.23) were inoculated with Lactococcus lactis ssp. cremoris starter culture (8 log cfu/mL) with or without Lactobacillus curvatus or Pediococci acidilactici as adjunct cultures (2 log cfu/mL). Cheddar cheeses were aged at 7.2 or 10°C for 168 d. The raw milk silo, ultrafiltration unit, cheddaring belt, and cheese tower had NSLAB biofilms ranging from 2 to 4 log cfu/100 cm². The population of Lb. curvatus reached 8 log cfu/g, whereas P. acidilactici reached 7 log cfu/g of experimental Cheddar cheese in 14 d. Higher NSLAB counts were observed in the first 14 d of aging in cheese stored at 10°C compared with that stored at 7.2°C. However, microbial counts decreased more quickly in Cheddar cheeses aged at 10°C compared with 7.2°C after 28 d. In cheeses without specific adjunct cultures (Lb. curvatus or P. acidilactici), calcium lactate crystals were not observed within 168 d. However, crystals were observed after only 56 d in cheeses containing Lb. curvatus, which also had increased concentration of d(−)-lactic acid compared with control cheeses. Our research shows that low levels of contamination with certain NSLAB can result in calcium lactate crystals, regardless of lactose:protein ratio.     

Mexican Queso Chihuahua: rheology of fresh cheese†

Rheological properties of fresh Queso Chihuahua made from raw milk (RM) or pasteurized milk (PM) were characterized using texture profile, torsion and small amplitude oscillatory shear analyses. Although the rheological properties varied significantly among the different brands with overlapping ranges for the individual RM and PM cheese brands, overall the PM cheeses were harder, chewier and more cohesive but had lower viscoelastic values than the RM cheeses. Establishing the rheological properties of Mexican Queso Chihuahua increases our understanding of the quality traits of Hispanic‐style cheeses and provides a foundation for maintaining the traditional texture of the cheese.     

Volatile fraction and sensory characteristics of Manchego cheese. 1. Comparison of raw and pasteurized milk cheese

Manchego cheese can be manufactured from raw or pasteurized ewes' milk. An automatic purge and trap apparatus, coupled to a GC-MS was used to isolate. identify and compare the relative amounts of the volatile components of raw and pasteurized Manchego cheese during ripening. The majority of volatile compounds were more abundant in raw milk (RM) cheeses than in pasteurized milk (PM) cheeses. Alcohols and esters predominated in the profile of RM Manchego cheese, while methyl-ketones and 2,3-butanedione were quantitatively important in PM cheeses. Branched chain alcohols were much more abundant in RM cheeses. The discriminant analysis separated 100% samples into RM or PM cheeses by using only 16 volatile compounds. Aroma intensity was correlated with esters, branched chain aldehydes and branched chain alcohols in RM cheeses, and with esters, branched chain aldehydes, 2-methyl ketones and 2-alkanols in PM cheeses. Diacetyl was positively correlated with the aroma attribute 'toasted' and negatively correlated with aroma quality in PM cheeses.     

Lipolysis in cheddar cheese made from raw, thermized, and pasteurized milks

The evolution of free fatty acids (FFA) was monitored over 168 d of ripening in Cheddar cheeses manufactured from good quality raw milk (RM), thermized milk (TM; 65°C × 15 s), and pasteurized milk (PM; 72°C × 15 s). Heat treatment of the milk reduced the level and diversity of raw milk microflora and extensively or wholly inactivated lipoprotein lipase (LPL) activity. Indigenous milk enzymes or proteases from RM microflora influenced secondary proteolysis in TM and RM cheeses. Differences in FFA in the RM, TM, and PM influenced the levels of FFA in the subsequent cheeses at 1 d, despite significant losses of FFA to the whey during manufacture. Starter esterases appear to be the main contributors of lipolysis in all cheeses, with LPL contributing during production and ripening in RM and, to a lesser extent, in TM cheeses. Indigenous milk microflora and nonstarter lactic acid bacteria appear to have a minor contribution to lipolysis particularly in PM cheeses. Lipolytic activity of starter esterases, LPL, and indigenous raw milk microflora appeared to be limited by substrate accessibility or environmental conditions over ripening.     

Occurrence of foodborne pathogens and characterization of Staphylococcus aureus in cheese produced on farm-dairies

The objective of this study was to address knowledge gaps identified in an earlier risk assessment of Staphylococcus aureus and raw milk cheese. A survey of fresh and short-time ripened cheeses produced on farm-dairies in Sweden was conducted to investigate the occurrence and levels of S. aureus, Listeria monocytogenes and Escherichia coli, to characterize S. aureus isolates with special emphasis on enterotoxin genes, antibiotic resistance, bio-typing and genetic variation, and to collect information related to production practices. In general, the hygienic quality of farm-dairy cheeses appeared to be of an acceptable microbiological quality, e.g. L. monocytogenes and staphylococcal enterotoxin were not detected in cheese samples. However, E. coli and enterotoxigenic S. aureus were frequently found in raw milk cheeses and sometimes at levels that are of concern, especially in fresh cheese. Interestingly, levels in raw milk fresh cheese were significantly lower when starter cultures were used. Up to five S. aureus colonies per cheese, if possible, were characterized and about 70% of isolates carried one or more enterotoxin genes, most common were sec and sea. The Ovine biotype (73%) was most common among isolates from goat milk cheese and the Human biotype (60%) from cow milk cheese. Of all isolates, 39% showed decreased susceptibility to penicillin, but the proportion of isolates from cows' cheese (66%) compared to isolates from goats' cheese (27%) was significantly higher. S. aureus isolates with different properties were detected in cheese from the same farm and, sometimes even the same cheese. Isolates with the same pulsed-field gel electrophoresis (PFGE)-pattern were detected on geographically distant dairies. This indicates that multiple sources and routes of contamination are important. To improve the safety of these products efforts to raise awareness of the importance of hygiene barriers and raw milk quality as well as improved process control can be suggested, e.g. use of starter cultures and monitoring of fermentation with a pH-meter. For future safety assessments, a better understanding of factors determining toxin production in these cheeses is needed.     

Volatile composition and sensory properties of industrially produced Idiazabal cheese

The volatile composition and sensory properties of industrially produced Idiazabal cheeses made from ewes’ raw milk (RM) or pasteurised milk (PM) and with addition of different starter cultures were compared. Cheeses were analysed at 90 and 180 d of ripening. Acids were the major volatile compounds in RM cheeses. Methyl ketones were the major volatile compounds in PM cheeses at 90 ripening days. However, the content of acids strongly increased with ripening whereas the content of ketones decreased in PM cheeses. The concentration of esters was higher in RM cheeses than in PM cheeses. No differences were found in the content of alcohols. Most aldehydes, hydrocarbons, terpenes and furans identified were minor volatile compounds in both RM and PM cheeses. In RM cheeses, characteristic sensory attributes for the aroma of Idiazabal cheese were present at 3 months, whereas in PM cheeses those desirable sensory attributes did not appear until 6 months of ripening.     

Short communication: Characterization of Shiga toxin 2-carrying bacteriophages induced from Shiga-toxigenic Escherichia coli isolated from Italian dairy products

Forty samples of raw milk cheese and 25 samples of raw milk itself were subjected to enrichment culture for Shiga-toxigenic Escherichia coli (STEC), and a single Shiga toxin 2- (Stx₂) positive strain was obtained from one of the cheese samples. Thus, aged cheeses in which the curd is subsequently heat treated (48°C) cannot be presumed to be STEC free. Infective Stx₂ bacteriophages were induced from 3 STEC strains isolated elsewhere from raw milk and 1 STEC strain from aged cheese sourced in Italy. Data on E. coli host range, morphology, genome size, and genetic variation determined by restriction fragment length polymorphism and multi-locus genotyping are presented. Although all 4 bacteriophages were found to be short-tailed Podoviridae, they exhibited considerable variation in both genome size and content. This extended to the Stx₂ genes themselves, whose sequences contained several point mutations, but these did not translate to amino acid substitutions.     

Ripening of Cheddar cheese made from blends of raw and pasteurised milk

Cheddar cheeses were made from pasteurised milk (P), raw milk (R) or pasteurised milk to which 10 (PR10), 5 (PR5) or 1 (PR1) % of raw milk had been added. Non-starter lactic acid bacteria (NSLAB) were not detectable in P cheese in the first month of ripening, at which stage PR1, PR5, PR10 and R cheeses had 10⁴, 10⁵, 10⁶ and 10⁷ cfu NSLAB g⁻¹, respectively. After ripening for 4 months, the number of NSLAB was 1–2 log cycles lower in P cheese than in all other cheeses. Urea–polyacrylamide gel electrophoretograms of water-soluble and insoluble fractions of cheeses and reverse-phase HPLC chromatograms of 70% (v/v) ethanol-soluble as well as -insoluble fractions of WSF were essentially similar in all cheeses. The concentration of amino acids were pro rata the number of NSLAB and were the highest in R cheese and the lowest in P cheese throughout ripening. Free fatty acids and most of the fatty acid esters in 4-month old cheeses were higher in PR1, PR5, PR10 and R cheeses than in P cheese. Commercial graders awarded the highest flavour scores to 4-month-old PR1 cheeses and the lowest to P or R cheese. An expert panel of sensory assessors awarded increasingly higher scores for fruity/sweet and pungent aroma as the level of raw milk increased. The trend for aroma intensity and perceived maturity was R>PR10>PP5>PR1>P. The NSLAB from raw milk appeared to influence the ripening and quality of Cheddar cheese.     

Survival of a five-strain cocktail of Escherichia coli O157:H7 during the 60-day aging period of cheddar cheese made from unpasteurized milk

The U.S. Food and Drug Administration Standard of Identity for Cheddar cheeses requires pasteurization of the milk, or as an alternative treatment, a minimum 60-day aging at > or =2 degrees C for cheeses made from unpasteurized milk, to reduce the number of viable pathogens that may be present to an acceptable risk. The objective of this study was to investigate the adequacy of the 60-day minimum aging to reduce the numbers of viable pathogens and evaluate milk subpasteurization heat treatment as a process to improve the safety of Cheddar cheeses made from unpasteurized milk. Cheddar cheese was made from unpasteurized milk inoculated with 10(1) to 10(5) CFU/ml of a five-strain cocktail of acid-tolerant Escherichia coli O157:H7. Samples were collected during the cheese manufacturing process. After pressing, the cheese blocks were packaged into plastic bags, vacuum sealed, and aged at 7 degrees C. After 1 week, the cheese blocks were cut into smaller-size uniform pieces and then vacuum sealed in clear plastic pouches. Samples were plated and enumerated for E. coli O157:H7. Populations of E. coli O157:H7 increased during the cheese-making operations. Population of E. coli O157:H7 in cheese aged for 60 and 120 days at 7 degrees C decreased less than 1 and 2 log, respectively. These studies confirm previous reports that show 60-day aging is inadequate to eliminate E. coli O157:H7 during cheese ripening. Subpasteurization heat-treatment runs were conducted at 148 degrees F (64.4 degrees C) for 17.5 s on milk inoculated with E. coli O157:H7 at 10(5) CFU/ml. These heat-treatment runs resulted in a 5-log E. coli O157: H7 reduction.     

Survey of raw milk cheeses for microbiological quality and prevalence of foodborne pathogens

Cheese may be manufactured in the United States using raw milk, provided the cheese is aged for at least 60 days at temperatures not less than 35 °F (1.7 °C). There is now increased concern among regulators regarding the safety of raw milk cheese due to the potential ability of foodborne pathogens to survive the manufacturing and aging processes. In this study, 41 raw milk cheeses were obtained from retail specialty shops, farmers’ markets, and on-line sources. The cheeses were then analyzed for the presence of Listeria monocytogenes, Salmonella, Escherichia coli O157:H7, Staphylococcus aureus, and Campylobacter. Aerobic plate counts (APC), coliform and yeast/mold counts were also performed. The results revealed that none of the enteric pathogens were detected in any of the samples tested. Five samples contained coliforms; two of those contained E. coli at less than 10² cfu/g. Three other cheese samples contained S. aureus. The APC and yeast-mold counts were within expected ranges. Based on the results obtained from these 41 raw milk cheeses, the 60-day aging rule for unpasteurized milk cheeses appears adequate for producing microbiologically safe products.     

Mexican Queso Chihuahua: functional properties of aging cheese

Queso Chihuahua, a semi-hard cheese manufactured from raw milk (RM) in northern Mexico, is being replaced by pasteurized milk (PM) versions because of food safety concerns and the desire for longer shelf life. In this study, the functional traits of authentic Mexican Queso Chihuahua made from RM or PM were characterized to identify sources of variation and to determine if pasteurization of the cheese milk resulted in changes to the functional properties. Two brands of RM cheese and 2 brands of PM cheese obtained in 3 seasons of the year from 4 manufacturers in Chihuahua, Mexico, were analyzed after 0, 4, 8, 12, and 16 wk of storage at 4°C. A color measurement spectrophotometer was used to collect color data before and after heating at 232°C for 5 min or 130°C for 75 min. Meltability was measured using the Schreiber Melt Test on samples heated to 232°C for 5 min. Sliceability (the force required to cut through a sample) was measured using a texture analyzer fitted with a wire cutter attachment. Proteolysis was tracked using sodium dodecyl sulfate-PAGE. Compared with PM cheeses, RM cheeses showed less browning upon heating, melted more at 232°C, and initially required a greater cutting force. With aging, cheeses increased in meltability, decreased in whiteness when measured before heating, and required less cutting force to slice. Seasonal variations in the cheesemilk had minimal or no effect on the functional properties. The differences in the functional properties can be attributed, in part, to the mixed microflora present in the RM cheeses compared with the more homogeneous microflora added during the manufacture of PM cheeses. The degree of proteolysis and subsequent integrity of the cheese matrix contribute to melt, slice, and color properties of the RM and PM cheeses. Understanding the functional properties of the authentic RM cheeses will help researchers and cheesemakers develop pasteurized versions that maintain the traditional traits desired in the cheeses.     

Short communication: Molecular characterization and antimicrobial resistance of pathogenic Escherichia coli isolated from raw milk and Minas Frescal cheeses in Brazil

The aim of this study was to quantify, identify, evaluate antimicrobial resistance, and characterize the virulence factors of enteropathogenic (EPEC), Shiga-toxigenic (STEC), and enterohemorrhagic (EHEC) Escherichia coli in raw milk (RM) and legal (LMFC) and illegal (IMFC) Minas Frescal cheeses in southern and northeast Brazil. Illegal cheeses are those made without official inspection service or sanitary surveillance. We evaluated samples of RM produced in Paraná (southern) and Maranhão (northeast) States, LMFC produced using pasteurized milk in inspected industries, and IMFC potentially produced with raw milk. Mean total coliform counts were 8.4 × 10⁴ cfu/mL for RM, 1.4 × 10⁷ cfu/mL for LMFC, and 2.9 × 10⁷ cfu/mL for IMFC. Mean E. coli counts were 2.4 × 10³ cfu/mL for RM, 1.9 × 10² cfu/mL for LMFC, and 1.1 × 10⁵ cfu/mL for IMFC. Among the 205 E. coli isolates from RM, 9.75% were identified as EPEC, mainly (90%) in samples from Paraná. Of the total isolates from the cheese samples, 97.4% (n = 111) came from IMFC, of which 1.8 and 2.7% were identified as EPEC and STEC, respectively; no EHEC was detected. The phylogenetic group A (60%) and typical EPEC (68%) predominated, which confirms the possible human origin of pathogenic isolates in RM and IMFC. Of these, 50% were resistant to at least one antibiotic, and streptomycin was the antimicrobial with the highest number (8) of EPEC and STEC resistant isolates. This study reports the first isolation of serogroup O28ac in Brazilian milk. We found no predominance of a specific serogroup of EPEC or STEC in milk or cheese or clonal isolates in the same sample, indicating different origins of the contamination in these products, presumably mostly related to poor hygienic handling.     

Impact of low concentration factor microfiltration on the composition and aging of Cheddar cheese

The effect of microfiltration (MF) on proteolysis, hardness, and flavor of Cheddar cheese during 6 mo of aging was determined. Raw skim milk was microfiltered two-fold in two cheese making trials. In trial 1, four vats of cheese were made in 1 d using unconcentrated milk (1X), 1.26X, 1.51X, and 1.82X concentration factors (CF). Casein-(CN)-to-fat ratio was constant among treatments. Proteolysis during cheese aging decreased with increasing CF due to either limitation of substrate availability for chymosin due to low moisture in the nonfat substance (MNFS), inhibition of chymosin activity by high molecular weight milk serum proteins, such as alpha2-macroglobulin, retained in the cheese or low residual chymosin in the cheese. Hardness of fresh cheese increased, and cheese flavor intensity decreased with increasing CF. In trial 2, the 1X and 1.8X CF were compared directly. Changes made in the cheese making procedure for the 1.8X CF (more chymosin and less cooking) increased the MNFS and made proteolysis during aging more comparable for the 1X and 1.8X cheeses. The significant difference in cheese hardness due to CF in trial 1 was eliminated in trial 2. In a triangle test, panelists could not differentiate between the 1X and 1.8X cheeses. Therefore, increasing chymosin and making the composition of the two cheeses more similar allowed production of aged Cheddar cheese from milk concentrated up to 1.8X by MF that was not perceived as different from aged Cheddar cheese produced without MF.     

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

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

Staphylococcus aureus and Listeria monocytogenes in Norwegian raw milk cheese production

The aim of this study was to survey the presence of Staphylococcus aureus and Listeria monocytogenes during the cheese making process in small-scale raw milk cheese production in Norway.The prevalence of S. aureus in bovine and caprine raw milk samples was 47.3% and 98.8%, respectively. An increase in contamination during the first 2-3 h resulted in a 73.6% prevalence of contamination in the bovine curd, and 23 out of 38 S. aureus-negative bovine milk samples gave rise to S. aureus-positive curds. The highest contamination levels of S. aureus were reached in both caprine and bovine cheese after 5-6 h (after the first pressing). There was no contamination of L. monocytogenes in caprine cheeses and only one (1.4%) contaminated bovine cheese.This work has increased our knowledge about S. aureus and L. monocytogenes contamination during the process of raw milk cheese production and gives an account of the hygiene status during the manufacture of Norwegian raw milk cheeses.     

Short communication: characterization of microflora in Mexican Chihuahua cheese

This work was performed to identify the bacterial species present in 10 Chihuahua cheeses obtained from commercial producers in Mexico using 16S rRNA gene analysis. As expected, some of the agar media initially used for isolation were not very selective, supporting the growth of several unrelated bacterial species. Sequence analysis identified potential pathogens, including Escherichia coli and Staphylococcus aureus, in all raw milk samples and 2 pasteurized milk samples. Streptococcus thermophilus and Lactococcus lactis ssp. lactis were identified in 9 and 6 samples, respectively, and would serve as acidifying agents during cheese production. Lactobacilli were identified in all cheeses, with the most prevalent being Lactobacillus plantarum identified in 7 raw milk and 1 pasteurized milk cheeses. Leuconostoc mesenteroides and Streptococcus macedonicus were identified in 4 raw milk cheeses and both were present in all pasteurized milk samples, suggesting that they may play a role in the development of traditional Chihuahua cheese attributes.     

Survival of Listeria monocytogenes introduced as a post-aging contaminant during storage of low-salt Cheddar cheese at 4, 10, and 21°C

Traditional aged Cheddar cheese does not support Listeria monocytogenes growth and, in fact, gradual inactivation of the organism occurs during storage due to intrinsic characteristics of Cheddar cheese, such as presence of starter cultures, salt content, and acidity. However, consuming high-salt (sodium) levels is a health concern and the dairy industry is responding by creating reduced-salt cheeses. The microbiological stability of low-salt cheese has not been well documented. This study examined the survival of L. monocytogenes in low-salt compared with regular-salt Cheddar cheese at 2 pH levels stored at 4, 10, and 21°C. Cheddar cheeses were formulated at 0.7% and 1.8% NaCl (wt/wt) with both low and high pH and aged for 10 wk, resulting in 4 treatments: 0.7% NaCl and pH 5.1 (low salt and low pH); 0.7% NaCl and pH 5.5 (low salt and high pH); 1.8% NaCl and pH 5.8 (standard salt and high pH); and 1.8% NaCl and pH 5.3 (standard salt and low pH). Each treatment was comminuted and inoculated with a 5-strain cocktail of L. monocytogenes at a target level of 3.5 log cfu/g, then divided and incubated at 4, 10, and 21°C. Survival or growth of L. monocytogenes was monitored for up to 90, 90, and 30 d, respectively. Listeria monocytogenes decreased by 0.14 to 1.48 log cfu/g in all treatments. At the end of incubation at a given temperature, no significant difference existed in L. monocytogenes survival between the low and standard salt treatments at either low or high pH. Listeria monocytogenes counts decreased gradually regardless of a continuous increase in pH (end pH of 5.3 to 6.9) of low-salt treatments at all study temperatures. This study demonstrated that post-aging inoculation of L. monocytogenes into low-salt (0.7%, wt/wt) Cheddar cheeses at an initial pH of 5.1 to 5.5 does not support growth at 4, 10, and 21°C up to 90, 90, and 30 d, respectively. As none of the treatments demonstrated more than a 1.5 log reduction in L. monocytogenes counts, the need for good sanitation practices to prevent post-manufacturing cross contamination remains.     

The manufacture of miniature Cheddar-type cheeses from milks with different fat globule size distributions

A novel 2-stage gravity separation scheme was developed for fractionation of raw, whole bovine milk into fractions enriched in small (SFG) or large (LFG) fat globules. The volume mean diameter of fat globules in SFG, LFG or control (CTRL) milk was 3.45, 4.68 and 3.58 microm, respectively. The maximum in storage modulus (index of firmness) decreased with increasing fat globule size for rennet-induced gels formed from SFG, LFG or CTRL milks. Miniature (20 g) Cheddar cheeses were manufactured using each of the 3 milks. There were no significant (P > 0.05) differences in the pH, moisture and fat in dry matter levels between cheeses made using any of the 3 milks, however, the fat content of the cheese made using SFG milk was approximately 1% lower than that of cheese made using LFG or CTRL milk in each of the 2 trials. Image analysis of confocal scanning laser micrographs of the cheeses illustrated that the star volume of fat globules in the cheeses decreased significantly (P < or = 0.05) as the size of fat globules in the milks used for cheesemaking was reduced. This indicates that it is possible to manipulate the size distribution of fat globules in Cheddar cheese by adjusting the fat globule size distribution of the milk used for cheese-making. The concentration of free fatty acids (FFA) increased in all cheeses during ripening. At 120 d of ripening, the concentration of FFA varied significantly (P < or = 0.05 and P < or = 0.001 for trials 1 and 2, respectively) with fat globule size, with cheeses made in trial 2 from LFG, SFG or CTRL milks having total FFA levels of 3391, 2820 and 2612 mg/kg cheese, respectively.