Variability of bacterial biofilms of the "tina" wood vats used in the ragusano cheese-making process

Ragusano cheese is a "protected denomination of origin" cheese made in the Hyblean region of Sicily from raw milk using traditional wooden tools, without starter. To explore the Ragusano bacterial ecosystem, molecular fingerprinting was conducted at different times during the ripening and biofilms from the wooden vats called "tinas" were investigated. Raw milks collected at two farm sites, one on the mountain and one at sea level, were processed to produce Ragusano cheese. Raw milk, curd before and after cooking, curd at stretching time (cheese 0 time), and cheese samples (4 and 7 months) were analyzed by PCR-temporal temperature gel electrophoresis (PCR-TTGE) and by classical enumeration microbiology. With the use of universal primers, PCR-TTGE revealed many differences between the raw milk profiles, but also notable common bands identified as Streptococcus thermophilus, Lactobacillus lactis, Lactobacillus delbrueckii, and Enterococcus faecium. After the stretching, TTGE profiles revealed three to five dominant species only through the entire process of ripening. In the biofilms of the two tinas used, one to five species were detected, S. thermophilus being predominant in both. Biofilms from five other tinas were also analyzed by PCR-TTGE, PCR-denaturating gradient gel electrophoresis, specific PCR tests, and sequencing, confirming the predominance of lactic acid bacteria (S. thermophilus, L. lactis, and L. delbrueckii subsp. lactis) and the presence of a few high-GC-content species, like coryneform bacteria. The spontaneous acidification of raw milks before and after contact with the five tinas was followed in two independent experiments. The lag period before acidification can be up to 5 h, depending on the raw milk and the specific tina, highlighting the complexity of this natural inoculation system.     

Breed of cow and herd productivity affect milk nutrient recovery in curd,and cheese yield,efficiency and daily production

Little is known about cheese-making efficiency at the individual cow level, so our objective was to study the effects of herd productivity, individual herd within productivity class and breed of cow within herd by producing, then analyzing, 508 model cheeses from the milk of 508 cows of six different breeds reared in 41 multi-breed herds classified into two productivity classes (high v. low). For each cow we obtained six milk composition traits; four milk nutrient (fat, protein, solids and energy) recovery traits (REC) in curd; three actual % cheese yield traits (%CY); two theoretical %CYs (fresh cheese and cheese solids) calculated from milk composition; two overall cheese-making efficiencies (% ratio of actual to theoretical %CYs); daily milk yield (dMY); and three actual daily cheese yield traits (dCY). The aforementioned phenotypes were analyzed using a mixed model which included the fixed effects of herd productivity, parity, days in milk (DIM) and breed; the random effects were the water bath, vat, herd and residual. Cows reared in high-productivity herds yielded more milk with higher nutrient contents and more cheese per day, had greater theoretical %CY, and lower cheese-making efficiency than low-productivity herds, but there were no differences between them in terms of REC traits. Individual herd within productivity class was an intermediate source of total variation in REC, %CY and efficiency traits (10.0% to 17.2%), and a major source of variation in milk yield and dCY traits (43.1% to 46.3%). Parity of cows was an important source of variation for productivity traits, whereas DIM affected almost all traits. Breed within herd greatly affected all traits. Holsteins produced more milk, but Brown Swiss cows produced milk with higher actual and theoretical %CYs and cheese-making efficiency, so that the two large-framed breeds had the same dCY. Compared with the two large-framed breeds, the small Jersey cows produced much less milk, but with greater actual and theoretical %CYs, similar efficiencies and a slightly lower dCY. Compared with the average of the specialized dairy breeds, the three dual-purpose breeds (Simmental and the local Rendena and Alpine Grey) had, on average, similar dMY, lower actual and theoretical %CY, similar fat and protein REC, and slightly greater cheese-making efficiency.     

Real-time visual/near-infrared analysis of milk-clotting parameters for industrial applications

The economical profitability of the dairy industry is based on the quality of the bulk milk collected in the farms, therefore it was based on the herd level rather than on the individual animals at real time. Udder infection and stage of lactation are directly related to the quality of milk produced on the herd level. However, improvement of milk quality requires testing each animal's milk separately and continuously. Recently, it was postulated that online equipment can estimate milk quality according to its clotting parameters, and thus result in better economical return for cheese making. This study further investigated the potential application of the AfiLab™ equipment to provide real-time analysis of milk-clotting parameters for cheese manufacture and cheese yield on quarter (1018) and individual cow (277) levels. Days in milk, lactose, log SCC and udder infection were found to have a significant effect on curd firmness and cheese properties and yield. The results clearly indicate that: (a) the parameter Afi-CF determined with the AfiLab™ is suitable for assessing milk quality for its clotting parameters, a value which is not provided by merely measuring fat and protein content on the gland and the cow levels; (b) bacterial type is the single major cause of reduced milk quality, with variations depending on the bacterial species; and (c) early and late lactation also had negative effects on milk-clotting parameters. Cheese made from the various milk samples that were determined by the Afilab™ to be of higher quality for cheese making resulted in higher yield and better texture, which were related mainly to the bacterial species and stage of lactation.     

Study of Enterobacteriaceae throughout the manufacturing and ripening of hard goats' cheese

The evolution of the counts and the species of Enterobacteriaceae as well as some physico-chemical parameters (pH, α_w and NaCl and moisture contents) during manufacturing and ripening of a hard Spanish goats' cheese of the Armada-Sobado variety were studied. Enterobacteriaceae (mean log counts 4.45 g^-1 in milk) increased 0.71–2.18 log units in curd and afterwards decreased until they disappeared after 2–4 weeks of ripening. This premature disappearance seems to be due to the decrease in α_w values and in moisture contents. However, the low pH values, reached from the beginning of the ripening process, could also contribute to this phenomenon.
The most abundant species in milk was Serratia liquefaciens (57.5% of isolates), followed by Morganella morganii (27.5%), Hafnia alvei (5%), Klebsiella oxytoca (5%) and Yersinia enterocolitica (5%). Yersinia enterocolitica was not subsequently isolated from either curd or in cheese. Hafnia alvei numbers increased in curd and in 1-week-old cheese where this micro-organism was the most abundant (47.5% and 75% of the isolates respectively). Escherichia coli , which was not isolated from milk, curd or 1-week-old cheese, was the predominant organism in 2-week-old cheese (57.8% of isolates). This confirms the finding of other authors who have shown that it is one of the most resistant species in ripening cheeses.     

Winter feeding systems and dairy cow breed have an impact on milk composition and flavour of two Protected Designation of Origin French cheeses

This study investigates the effects of two feeding systems and two dairy cow breeds on milk yield and composition, physical and sensorial properties of Camembert and Pont-l’Evêque cheeses. The experiment consisted of a 2 × 2 factorial arrangement of treatments. A low energy grass diet with only 15% of concentrate (LowGS) was compared with a high-energy maize silage diet with 30% concentrate (HighMS). Thirty-four Holstein (Ho) and 34 Normande (No) cows in early lactation were assigned to one of two feeding systems for a 6-week period. Cows on the LowGS feeding system had lower milk yield, fat and protein content. In both feeding systems, No cows had lower milk yields but higher milk protein contents than Ho cows. The LowGS feeding system altered milk fatty acid (FA) composition by reducing saturated FA. Breed had only a small impact on milk FA. Concerning milk coagulating properties, only the firmness was reduced by the LowGS feeding and the Ho breed. The effects of breed and feeding system on the protein content of cheeses were more marked in Camembert cheese than in Pont-l’Evêque cheese. However, the Camembert cheese from Ho-LowGS was, in fact, characterized especially by lower protein content. LowGS feeding system and No breed produced more yellow cheeses. Feeding systems had limited effects on the firmness of Camembert and Pont-l’Evêque cheeses measured by penetrometry. In sensory analysis, Ho breed and LowGS feeding produced a Camembert cheese with a more melting texture in the mouth due to the increase of spreadability index and of proteolysis. The type of cheese also had an influence: the effects were more important on Camembert cheese than on Pont-l’Evêque cheese. Only the Ho-LowGS treatment produced a very specific Camembert cheese different from the others. The feeding systems and breed of dairy cow have no determinant effect on PDO (protected designation of origin) Camembert and Pont-l’Evêque cheeses, especially regarding taste. In this kind of trial, despite the effects of feeding systems and breed on milk composition, the role of cheese ripening and microbiology appears to be of considerable importance.     

Milk composition in the eastern quoll, Dasyurus viverrinus (Marsupialia:Dasyuridae)

The milk constituents of Dasyurus viverrinus, a carnivorous marsupial, exhibited major quantitative and qualitative changes during the course of lactation. The milk produced in the early stages of lactation was dilute, about 13-16% (w/w) solids before 3 weeks with carbohydrate representing the major fraction. In the latter stages of lactation the milk was concentrated, around 30% solids, and lipid was the predominant fraction. Palmitic acid was the major fatty acid present in early-stage milk but oleic acid became predominant in milk after 10 weeks post-partum. The changes in milk composition in D. viverrinus were similar to those described for the milks of herbivorous marsupials which therefore suggests that this pattern may be uniform throughout the Marsupialia.     

Once-a-day milking of Holstein and Montbéliarde cows for 7 weeks in mid-lactation

This study aimed at comparing the effects of once-a-day (OAD) milking during the descending phase of lactation between cows from the two most common breeds in France (Holstein and Montbéliarde). This study was carried on during two successive summers on a total of 50 Holstein and 38 Montbéliarde cows. During 7 weeks, half of the cows from each breed was milked OAD while the other half was milked twice a day (TAD). The animals were also followed for the next 3 or 5 weeks when they were all milked TAD, to check for any residual effect of OAD milking. The behaviour of OAD cows was observed around milking time. The incidence of diseases, the main performance variables (milk production, milk flow rate, live weight and body condition score), the detailed composition of milk (fat, protein, lactose, somatic cells, minerals, pH, free fatty acid (FFA), nitrogen fractions and enzymes) and some technological variables (clotting time and curd firmness) were measured on all cows.Some signs of disturbance were observed in the OAD cows at the time when milking was omitted: some cows mooed, some went close to the exit of the paddock, some leaked milk prior to milking. However, these signs disappeared after 2 days. After the experimental period, the live weight and the body condition score of TAD and OAD cows did not differ significantly. OAD cows produced 4.5 kg/day less than TAD (P < 0.001), this being more marked in Holstein (5.7 kg/day, P < 0.001) than Montbéliarde (3.3 kg/day, P < 0.001) cows. The milk contents of fat, whey protein, casein, total protein and phosphorus, and its plasminogen activity, were higher with OAD cows while lactose and FFA contents, and lipoprotein lipase activity were lower, with no interaction observed with breed. During the subsequent 3 weeks, when all cows were again milked TAD, OAD cows still produced 1.7 kg/day less milk (P < 0.01) with slightly higher fat and protein content.OAD milking for 7 weeks during the descending phase of lactation decreased milk production but increased milk content of most components, with a low residual effect. Montbéliarde cows were less affected by OAD milking than Holstein cows.     

Popular ovine and caprine fermented milks

Ovine and caprine milk are widely produced in semi-arid countries, and mainly utilised for milk consumption and the manufacture of a wide range of cheeses, fermented milk products (e.g. liquid, viscous, concentrated and dried) and to a lesser degree milk powder. The primary aim of this review is to provide a critical analysis of the main components of milk from different ovine and caprine breeds for the manufacture of fermented milk products, i.e. traditional and industrial. This paper reviews the latest scientific developments in fermented milks, including probiotic products, and recommendations to improve the characteristics of the final product(s). Although ovine milk is successfully used to manufacture many fermented milk products, the solids content of milk from certain breeds of caprine needs to be fortified to enhance the protein content and modify the processing conditions to ensure products acceptable to the consumer.     

Yeast populations associated with the artisanal cheese produced in the region of Serra da Canastra, Brazil

The aim of this work was to describe the yeast populations present during the manufacturing of Minas cheese of the region of Serra da Canastra, Minas Gerais state, Brazil. Canastra cheese is produced from raw cow’s milk at the farmhouse level using artisanal procedures and natural whey cultures as starters. Samples from 10 farms were studied, and they included: raw milk, natural starter, cheese curd before salting and cheese after 5 days of ripening. The most frequent yeasts in whey, curd and cheese were Debaryomyces hansenii, Kluyveromyces lactis, Kodamaea ohmeri and Torulaspora delbrueckii. Many yeast isolates were able to produce proteases, lipases and β-galactosidades. Production of these enzymes by yeasts in the cheese would contribute to the development of the characteristic flavor and smell during the ripening process.     

Association of Yersinia enterocolitica with the manufacture of cheese and occurrence in pasteurized milk.

Raw milk in southern Ontario frequently contains Yersinia enterocolitica. The potential for transmission of this organism by cheese manufactured from unpasteurized milk was evaluated by examination of milk and cheese curd samples from cheese manufacturing plants and finished cheddar and Italian cheeses. The incidence of Y. enterocolitica was lower in cheese curd samples (9.2%) than in raw milk (18.2%). Most of the curd samples showed a positive phosphatase test, indicating production from raw milk. One curd sample yielded Y. enterocolitica after 4 weeks of storage at 4 degrees C but was negative after 8 weeks. All samples of cheddar and Italian cheeses, most of which showed a positive phosphatase test, were negative for Y. enterocolitica. One out of 265 samples (0.4%) of pasteurized fluid dairy products contained Y. enterocolitica.     

Effects of restricted feeding of prepubertal ewe lambs on reproduction and lactation performances over two breeding seasons

The aim of this study was to determine the effects of restricted feeding before puberty on reproduction, lactation and offspring growth performance in replacement ewe lambs over two breeding seasons. At weaning, 41 Dorset ewe lambs were assigned to one of three diets: an ad libitum control diet with medium-quality forage (MQF; 13.3% crude protein (CP), 1.81 Mcal metabolizable energy per kg, 42.8% ADF; diet A-MQF); a restricted diet with the same forage as A but less feed concentrate (diet R-MQF); or a high-quality forage (HQF) diet (14.8% CP, 2.15 Mcal ME/kg, 34.7% ADF; diet F-HQF). The quantity of concentrate offered to the group R-MQF and F-HQF ewe lambs was adjusted to obtain 70% of the control ewe lambs' growth rate. The diets were offered for 75 days following weaning to cover the allometric phase of mammary gland development. Prepubertal restriction did not affect (P > 0.10) the gestation rate, number of lambs born or the body weight and body condition score of ewes at lambing or at the end of lactation. Ewes from groups R-MQF and F-HQF tended to produce more milk during their first lactation compared to those from group A-MQF (P = 0.07). During the second lactation, groups R-MQF and F-HQF had better standardized milk production than group A-MQF (P < 0.05), and group R-MQF produced more milk than group F-HQF (P < 0.05). Milk fat and protein content were not affected by treatments (P > 0.10) Fat and protein yield were affected by treatments only at the second lactation (P < 0.10 and P < 0.05, respectively). Lamb birth and weaning weights were not affected by prepubertal restriction of feeding in their mother (P > 0.10). However, the average daily gain of second breeding season lambs was higher for the R-MQF group than the F-HQF group (P < 0.05), and a similar trend was observed for total gain (P < 0.10). Restricted feeding before puberty does not impair future reproductive performance; however, it has a positive impact on lactation and on lambs' growth performance.     

Acceleration of cheese ripening

The characteristic aroma, flavour and texture of cheese develop during ripening of the cheese curd through the action of numerous enzymes derived from the cheese milk, the coagulant, starter and non-starter bacteria. Ripening is a slow and consequently an expensive process that is not fully predictable or controllable. Consequently, there are economic and possibly technological incentives to accelerate ripening. The principal methods by which this may be achieved are: an elevated ripening temperature, modified starters, exogenous enzymes and cheese slurries. The advantages, limitations, technical feasibility and commercial potential of these methods are discussed and compared.     

Heritability and repeatability of milk coagulation properties predicted by mid-infrared spectroscopy during routine data recording,and their relationships with milk yield and quality traits

The aim of this study was to estimate (co)variance components for milk coagulation properties (MCP) predicted by mid-infrared spectroscopy (MIRS) during routine milk recording, and to assess their relationships with yield and quality traits. A total of 63 470 milk samples from Holstein-Friesian cows were analyzed for MCP, pH and quality characteristics using MIRS. Casein to protein and protein to fat ratios were calculated from information obtained by MIRS. Records were collected across 1 year on 16 089 cows in 345 herds. The model used for genetic analysis included fixed effects of parity and stage of lactation, and random effects of herd-test-day, cow permanent environmental, animal additive genetic and residual. (Co)variance components were assessed in a Bayesian framework using the Gibbs Sampler. Estimates of heritabilities were consistent with those reported in the literature, being moderate for MCP (0.210 and 0.238 for rennet coagulation time (RCT) and curd firmness (a₃₀), respectively), milk contents (0.213 to 0.333) and pH (0.262), and low for somatic cell score (0.093) and yield traits (0.098 to 0.130). Repeatabilities were 0.391 and 0.434 for RCT and a₃₀, respectively, and genetic correlations were generally low, with estimates greater than 0.30 (in absolute value) only for a₃₀ with fat, protein and casein contents. Overall, results suggest that genetic evaluation for MCP predicted by MIRS is feasible at population level, and several repeated measures per cow during a lactation are required to estimate reliable breeding values for coagulation traits.     

Gelsolin expression in sheep milk somatic cells during lactation

The identification of genes involved in phenotypes related to milk quality is important for both economic and health aspects in livestock production. The aim of this study was to assess the level of gelsolin gene expression in two breeds of dairy sheep – Sarda and Gentile – with pronounced differences in quantitative and qualitative milk traits. Gelsolin, a type of actin-modulating proteins is involved in the processes of actin remodeling during cell growth and apoptosis; therefore a role of this protein in mammary changes during lactation was here hypothesized. Individual milk samples were collected three times during lactation from 26 ewes of the two breeds. The differential gene expression of gelsolin in the two breeds and the three lactation times was estimated by quantitative PCR on RNA extracted from milk somatic cells. Correlations of gelsolin gene expression with milk yield and quality and days of lactation were also estimated. The results showed that gelsolin gene expression was significantly higher in the Sarda compared to the Gentile at each lactation stage, in agreement with the longer lactation duration and the higher daily milk yield of the first breed. Significant correlations of gelsolin gene expression were found with milk fat content in Sarda breed (−0.46, P<0.05). Gelsolin expression analysis confirmed the link between gelsolin gene function and milk fat content of sheep.     

Microbiological characteristics of anevato: a traditional greek cheese

Nine batches of Anevato, raw goat milk cheese, were examined throughout a 60 day storage time at three different periods within the lactation season of the goat. High mean log counts per gram of cheese for aerobic bacteria (7·92–9·56), lactic acid bacteria (7·78–9·32), Gram-negative organisms 5·64–9·67), psychrotrophs (7·90–11·79) and proteolytic bacteria (7·57–9·36) were found. Enterobacteriaceae, coliforms and yeasts were considerably lower. Enterobacteriaceae and coliforms in the curd of cheese made in May were lower by approximately 3·0 log₁₀ cfu g⁻¹ than counts in curd made in January, and were lower by about 2·5 log₁₀ cfu g⁻¹ than those in cheese made in March. This coincided with lower pH and higher counts of lactic acid bacteria in cheese made in March and May. Yeast populations were affected by the season and were higher in May than March and/or January. Lactococci dominated in the cheese until 15 days, but lactobacilli became predominant after 30 days. Lactococcus lactis was the most abundant species of lactic acid bacteria found in Anevato cheese. Results suggest the need for improving milk quality and/or using heat-treated milk to produce Anevato cheese; the use of L. lactis as a starter would possibly eliminate or suppress the growth of undesirable organisms.     

Behavior of Salmonella typhimurium During Manufacture and Curing of Cheddar Cheese

Nine vats of stirred-curd granular cheddar cheese were made with whole milk contaminated with Salmonella typhimurium after pasteurization. Enumeration of salmonellae by the most probable number technique during manufacture and curing showed that these organisms multiplied rapidly during manufacture until the curd was salted. Thereafter and throughout the curing period, the salmonellae declined in number at a rate dependent on the temperature of curing. Evidence is presented indicating that the production of volatile fatty acids in the curd during curing may be responsible for this decline.     

Effects of aging on functional properties of caprine milk made into Cheddar- and Colby-like cheeses

The effects of cheese milk obtained at three times during lactation (weeks 4–5, 12–15, and 21–23) and cheese storage (up to 16 or 24 weeks) on meltability, sliceability, and color changes upon heating (232 °C for 5 min, high baking temperature, HT, or 130 °C for 75 min, low baking temperature, LT) of caprine milk cheeses were evaluated. The cheeses were manufactured from milk from Alpine goats and based on the procedures of Cheddar and Colby cheese manufacture. In Cheddar-like cheese, the sliceability (force required to slice sample) was at its highest when the cheese was made with milk from weeks 12–15 into lactation. Color change was variable although it tended to be lowest in cheese made at weeks 4–5 into lactation. In Colby-like cheeses, meltability was at its highest and sliceability was very poor (after 8 weeks of aging) when made with milk obtained later in lactation. Color changes were variable at the two different baking temperatures. As expected during aging, the meltability of the cheeses increased and the force required to slice the cheeses decreased with the significant changes occurring within the first 16 weeks for Cheddar-like and the first 8 weeks for Colby-like cheeses. The color changes upon heating were variable for aged Cheddar-like cheeses and did not change significantly for aged Colby-like cheeses. Color changes were highly correlated with proteolysis occurring during storage. Cheese milk obtained at different times of lactation and aging of the cheese impact the functional properties of caprine milk cheeses and will affect their optimal utilization.     

Preliminary characterization of microflora of Comté cheese

The evolution of the microflora of three Comté cheeses made in duplicate with raw milk from three different sources was followed during ripening. The same starter was used with each type of milk. The comparison of the cheeses did not reveal any significant difference in the development of the microflora. Starter lactic acid bacteria ( Streptococcus thermophilus and Lactobacillus helveticus) , which are added at the beginning of manufacture, decreased quickly in the first stages of ripening supporting the hypothesis of cell autolysis. Other micro-organisms, i.e. homofermentative and heterofermentative lactobacilli ( Lact. delbrueckii ssp. lactis , Lact. paracasei ssp. paracasei , Lact. rhamnosus and Lact. fermentum ), pediococci, enterococci and propionibacteria grew in cheese from small numbers in fresh curd. The characterization of Strep. thermophilus by pulsed-field gel electrophoresis showed that wild strains were also able to grow in the curd. The values for the genome size of 11 Strep. thermophilus strains determined in this investigation were in the range of 1·8–2·3 Mbp. The potential role of starter and raw milk microflora in cheese flavour development was considered.     

Does the milk of callitrichid monkeys differ from that of larger anthropoids?

The generalization that anthropoid primates produce dilute milks that are low in protein and energy is based primarily on data from large monkeys of the families Cebidae and Cercopithecidae, as well as humans. The marmosets and tamarins (Callitrichidae) are not only much smaller in body size, but also typically raise multiple offspring during a relatively brief lactation. We hypothesized that selection for small body size and high reproductive rate might favor secretion of milk of higher energy and protein concentrations. To test this hypothesis, 46 milk samples collected from 10 common marmosets (Callithrix jacchus, ca. 350 g) were assayed for dry matter (DM), crude protein (CP), fat, and sugar; and gross energy (GE) was calculated from these constituents. We also assayed five samples collected from three golden lion tamarins (Leontopithecus rosalia, ca. 700 g) and six samples collected from a single pygmy marmoset (Cebuella pygmaea, ca. 150 g) over two lactation periods. All samples were collected between days 10 and 57 post partum, representing mid lactation for these species. The milks of these three species were similar, containing 14.0%, 16.1%, and 13.7% DM; 2.7%, 2.6%, and 2.9% CP; 3.6%, 5.2%, and 3.7% fat; 7.4%, 7.2%, and 7.8% sugar; and 0.76, 0.90, and 0.82 kcal/g for common marmosets, golden lion tamarins, and the pygmy marmoset, respectively. These species produced milks with energy values that were within the range reported for large anthropoids, albeit with slightly higher protein concentration. However, milk composition did vary substantially among individual common marmoset females, especially in the proportion of milk energy derived from fat. In contrast, CP as expressed as a percent of GE was remarkably constant among common marmoset females. Callitrichid milk appeared to be similar to that of larger anthropoid primates in GE, but was higher in CP and in the proportion of GE from CP. However, the small sample sizes for the golden lion tamarin and the pygmy marmoset, and the wide variation in milk composition found among common marmoset females cautions against definitively characterizing the milks of callitrichids from these data.     

Investigating the relationship between raw milk bacterial composition, as described by intergenic transcribed spacer–PCR fingerprinting, and pasture altitude

Aims:  To assess the bacterial biodiversity level in bovine raw milk used to produce Fontina, a Protected Designation of Origin cheese manufactured at high-altitude pastures and in valleys of Valle d'Aosta region (North-western Italian Alps) without any starters. To study the relation between microbial composition and pasture altitude, in order to distinguish high-altitude milk against valley and lowland milk.
Methods and Results:  The microflora from milks sampled at different alpine pasture, valley and lowland farms were fingerprinted by PCR of the 16S–23S intergenic transcribed spacers (ITS-PCR). The resulting band patterns were analysed by generalized multivariate statistical techniques to handle discrete (band presence–absence) and continuous (altitude) information. The fingerprints featured numerous bands and marked variability indicating complex, differentiated bacterial communities. Alpine pasture milks were distinguished from lowland ones by cluster analysis, while this technique less clearly discriminated alpine pasture and valley samples. Generalized principal component analysis and clustering-after-ordination enabled a more effective distinction of alpine pasture, valley and lowland samples.
Conclusions:  Alpine raw milks for Fontina production contain highly diverse bacterial communities, the composition of which is related to the altitude of the pasture where milk was produced.
Significance and Impact of the Study:  This research may provide analytical support to the important issue represented by the authentication of the geographical origin of alpine milk productions.