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Cheese is vulnerable to contamination with mycotoxins, particularly ochratoxin A (OTA) and aflatoxin M1 (AFM1). This study aims to develop and validate an analytical method for the detection and quantification of OTA and AFM1 in cheese and to assess their prevalence and associated risks. A liquid chromatography–tandem mass spectrometry (LC-MS/MS) method was validated for detecting these mycotoxins in 41 cheese samples, including firm-ripened, spreadable, and plant-based alternatives. The results showed that OTA was detected exclusively in grated Parmigiano Reggiano cheese, while AFM1 was found in both Parmigiano Reggiano and Pecorino cheeses. This study goes beyond analytical method development by providing a preliminary exposure assessment and risk characterization for OTA and AFM1 in cheese, bridging the gap between analytical chemistry and public health implications. This study identified potential health risks associated with OTA, particularly for children and adolescents categorized as high consumers of Parmigiano Reggiano cheese. The findings underscore the need for monitoring of OTA and AFM1 in cheese and further research to establish regulatory limits for these contaminants. [ABSTRACT FROM AUTHOR]

The aims of this proof of principle study were to compare two different chemometric approaches using a Bayesian method, Partial Least Square (PLS) and PLS-discriminant analysis (DA), for the prediction of the chemical composition and texture properties of the Grana Padano (GP) and Parmigiano Reggiano (PR) PDO cheeses by using NIR and Raman spectra and quantify their ability to distinguish between the two PDO and among their ripening periods. For each dairy chain consortium, 9 cheese samples from 3 dairy industries were collected for a total of 18 cheese samples. Three seasoning times were chosen for each dairy industry: 12, 20, and 36 months for GP and 12, 24, and 36 months for PR. A portable NIR instrument (spectral range: 950–1,650 nm) was used on 3 selected spots on the paste of each cheese sample, for a total of 54 spectra collected. An Alpha300 R confocal Raman microscope was used to collect 10 individual spectra for each cheese sample in each spot for a total of 540 Raman spectra collected. After the detection of eventual outliers, the spectra were also concatenated together (NIR + Raman). All the cheese samples were assessed in terms of chemical composition and texture properties following the official reference methods. A Bayesian approach and PLS-DA were applied to the NIR, Raman, and fused spectra to predict the PDO type and seasoning time. The PLS-DA reached the best performances, with 100% correctly identified PDO type using Raman only. The fusion of the data improved the results in 60% of the cases with the Bayesian and of 40% with the PLS-DA approach. A Bayesian approach and a PLS procedure were applied to the NIR, Raman, and fused spectra to predict the chemical composition of the cheese samples and their texture properties. In this case, the best performance in validation was reached with the Bayesian method on Raman spectra for fat (R2VAL = 0.74). The fusion of the data was not always helpful in improving the prediction accuracy. Given the limitations associated with our sample set, future studies will expand the sample size and incorporate diverse PDO cheeses.

Objective The aim was to study the influence of cooling milk at 9°C at the farm versus keeping it at 20°C on Parmigiano Reggiano cheese lipolysis. Methods A total of six cheesemaking trials (3 in winter and 3 in summer) were performed. In each trial, milk was divided continuously into two identical aliquots, one of which was kept at 9°C (MC9) and the other at 20°C (MC20). For each trial and milk temperature, vat milk (V-milk) and the resulting 21 month ripened cheese were analysed. Results Fat and dry matter and fat/casein ratio were lower in MC9 V-milk (p≤0.05) than in MC20. Total bacteria, mesophilic lactic acid and psychrotrophic and lipolytic bacteria showed significant differences (p≤0.05) between the two V-milks. Regarding cheese, fat content resulted lower and crude protein higher (p≤0.05) both in outer (OZ) and in inner zone (IZ) of the MC9 cheese wheels. Concerning total fatty acids, the MC9 OZ had a lower concentration of butyric, capric (p≤0.05) and medium chain fatty acids (p≤0.05), while the MC9 IZ had lower content of butyric (p≤0.05), caproic (p≤0.01) and short chain fatty acids (p≤0.05). The levels of short chain and medium chain free fatty acids (p≤0.05) were lower and that of long chain fatty acids (p≤0.05) was higher in MC9 OZ cheese. The principal component analysis of total and free fatty acids resulted in a clear separation among samples by seasons, whereas slight differences were observed between the two different milk temperatures. Conclusion Storing milk at 9°C at the herd affects the chemical composition of Parmigiano Reggiano, with repercussion on lipolysis. However, the changes are not very relevant, and since the cheese can present a high variability among the different cheese factories, such changes should be considered within the “normal variations” of Parmigiano Reggiano chemical characteristics.

In recent years, due to the significant increase in hypertension, peptides which are able to reduce blood pressure have gained special interest by scientific research and food industry. Several bioactive peptides with ascertained ACE-inhibitory activity have been found in Parmigiano Reggiano (PR) cheese and/or mixtures deriving from its digestion in vitro, and this may be predictive of its potential antihypertensive effect in vivo. This study investigated the long-term effect of feeding (PR) cheese on blood pressure (BP) of spontaneously hypertensive rats (SHRs). A total of 30 male SHRs, 13 weeks old, were subdivided into 6 groups balanced for body weight and BP, to receive daily dietary supplementation with: 0.1–0.2–0.4–0.6 g PR/rat, captopril, and water. Systolic and diastolic BP were recorded every two weeks, for 10 weeks. Blood samples were collected at the end of the trial. Dietary integration with PR led to a transitory reduction in rats’ pressure in the first 35 days of treatment and pressure decreased in a dose-dependent manner. In the second part of the study, the beneficial effect of PR antihypertensive peptides may have been masked and reduced by the increase in BP of rats linked to the rise in age of animals. No PR derived peptides were detected in rats’ serum. Highlights: Parmigiano Reggiano (PR) cheese led to a transitory reduction in rats’ pressure in the first 35 days of treatment. This effect was PR dose dependent. The highest amounts of PR tested did not increase both systolic and diastolic blood pressures of hypertensive rats.

Fermentation is a promising solution to valorize cheese whey, the main by-product of the dairy industry. In Parmigiano Reggiano cheese production, natural whey starter (NWS), an undefined community of thermophilic lactic acid bacteria, is obtained from the previous day residual whey through incubation at gradually decreasing temperature after curd cooking. The aim of this study was to investigate the effect of fermentation regime (spontaneous (S) and NWS-inoculated (I-NWS)) on biofunctionalities and release of bioactive peptides during whey fermentation. In S and I-NWS trials proteolysis reached a peak after 24 h, which corresponded to the drop out in pH and the maximum increase in lactic acid. Biological activities increased as a function of fermentation time. NWS inoculum positively affected antioxidant activity, whilst S overcame I-NWS in angiotensin-converting enzyme (ACE) and DPP-IV (dipeptidyl peptidase IV) inhibitory activities. Peptidomics revealed more than 400 peptides, mainly derived from β-casein, κ-casein, and α-lactalbumin. Among them, 49 were bioactive and 21 were ACE-inhibitors. Semi-quantitative analysis strongly correlated ACE-inhibitory activity with the sum of the peptide abundance of ACE-inhibitory peptides. In both samples, lactotripeptide isoleucine-proline-proline (IPP) was higher than valine-proline-proline (VPP), with the highest content in S after 24 h of fermentation. In conclusion, we demonstrated the ability of whey endogenous microbiota and NWS to extensively hydrolyze whey proteins, promoting the release of bioactive peptides and improving protein digestibility. [ABSTRACT FROM AUTHOR]

Objective: The aim was to study the influence of cooling milk at 9°C at the farm versus keeping it at 20°C on Parmigiano Reggiano cheese lipolysis. Methods: A total of six cheesemaking trials (3 in winter and 3 in summer) were performed. In each trial, milk was divided continuously into two identical aliquots, one of which was kept at 9°C (MC9) and the other at 20°C (MC20). For each trial and milk temperature, vat milk (V-milk) and the resulting 21 month ripened cheese were analysed. Results: Fat and dry matter and fat/casein ratio were lower in MC9 V-milk (p≤0.05) than in MC20. Total bacteria, mesophilic lactic acid and psychrotrophic and lipolytic bacteria showed significant differences (p≤0.05) between the two V-milks. Regarding cheese, fat content resulted lower and crude protein higher (p≤0.05) both in outer (OZ) and in inner zone (IZ) of the MC9 cheese wheels. Concerning total fatty acids, the MC9 OZ had a lower concentration of butyric, capric (p≤0.05) and medium chain fatty acids (p≤0.05), while the MC9 IZ had lower content of butyric (p≤0.05), caproic (p≤0.01) and short chain fatty acids (p≤0.05). The levels of short chain and medium chain free fatty acids (p≤0.05) were lower and that of long chain fatty acids (p≤0.05) was higher in MC9 OZ cheese. The principal component analysis of total and free fatty acids resulted in a clear separation among samples by seasons, whereas slight differences were observed between the two different milk temperatures. Conclusion: Storing milk at 9°C at the herd affects the chemical composition of Parmigiano Reggiano, with repercussion on lipolysis. However, the changes are not very relevant, and since the cheese can present a high variability among the different cheese factories, such changes should be considered within the “normal variations” of Parmigiano Reggiano chemical characteristics. [ABSTRACT FROM AUTHOR]

Time of ripening has a strong impact on shaping the valuable and recognizable characteristics of long-ripened types of cheese such as Parmigiano Reggiano (PR) due to the interrelationship between microbiota and proteolysis that occurs during ripening. The derived peptide profile is linked to cheese quality and represents the canvas for enzymes upon digestion, which could be responsible for the release of potentially bioactive peptides (BPs). In this study, we aimed at investigating the presence of BP in 72 PR cheese samples of different ripening times, from curd to 24 months of ripening, produced in six different dairies, and following their fate after simulated gastrointestinal digestion. A small number of peptide sequences sharing 100% similarity with known antimicrobial, antioxidant, and ACE-inhibitor sequences were found in PR cheeses, while a higher number of potential BPs were found after their simulated gastrointestinal digestion, in different amounts according to ripening time. Taking advantage of the complex organization of the sampling plan, we were able to follow the fate of peptides considered quality drivers during cheese ripening to their release as functional compounds upon digestion. [ABSTRACT FROM AUTHOR]

Time of ripening has a strong impact on shaping the valuable and recognizable characteristics of long-ripened types of cheese such as Parmigiano Reggiano (PR) due to the interrelationship between microbiota and proteolysis that occurs during ripening. The derived peptide profile is linked to cheese quality and represents the canvas for enzymes upon digestion, which could be responsible for the release of potentially bioactive peptides (BPs). In this study, we aimed at investigating the presence of BP in 72 PR cheese samples of different ripening times, from curd to 24 months of ripening, produced in six different dairies, and following their fate after simulated gastrointestinal digestion. A small number of peptide sequences sharing 100% similarity with known antimicrobial, antioxidant, and ACE-inhibitor sequences were found in PR cheeses, while a higher number of potential BPs were found after their simulated gastrointestinal digestion, in different amounts according to ripening time. Taking advantage of the complex organization of the sampling plan, we were able to follow the fate of peptides considered quality drivers during cheese ripening to their release as functional compounds upon digestion.

In recent years, due to the significant increase in hypertension, peptides which are able to reduce blood pressure have gained special interest by scientific research and food industry. Several bioactive peptides with ascertained ACE-inhibitory activity have been found in Parmigiano Reggiano (PR) cheese and/or mixtures deriving from its digestion in vitro, and this may be predictive of its potential antihypertensive effect in vivo. This study investigated the long-term effect of feeding (PR) cheese on blood pressure (BP) of spontaneously hypertensive rats (SHRs). A total of 30 male SHRs, 13 weeks old, were subdivided into 6 groups balanced for body weight and BP, to receive daily dietary supplementation with: 0.1–0.2–0.4–0.6 g PR/rat, captopril, and water. Systolic and diastolic BP were recorded every two weeks, for 10 weeks. Blood samples were collected at the end of the trial. Dietary integration with PR led to a transitory reduction in rats' pressure in the first 35 days of treatment and pressure decreased in a dose-dependent manner. In the second part of the study, the beneficial effect of PR antihypertensive peptides may have been masked and reduced by the increase in BP of rats linked to the rise in age of animals. No PR derived peptides were detected in rats' serum. Highlights: Parmigiano Reggiano (PR) cheese led to a transitory reduction in rats' pressure in the first 35 days of treatment. This effect was PR dose dependent. The highest amounts of PR tested did not increase both systolic and diastolic blood pressures of hypertensive rats. [ABSTRACT FROM AUTHOR]

This research communication depicts the quality and authenticity of grated Parmesan cheese sold in Brazil, with novel emphasis on ripening time. The sampling included all brands manufactured and packed at Brazilian dairy plants under federal inspection and sold in Londrina between June and September 2017. A total of 24 samples were analyzed (3 batches of 8 brands) for microbiological and compositional characteristics, including urea-PAGE electrophoresis for the quantitative determination of the milk proteins. About 80% of the samples were insufficiently ripened and 32% with low-fat content. Faced with these economically motivated food frauds, the authenticity of grated Parmesan in Brazil may be under threat. It is high recommended that future surveys and regulatory agencies go further than microbiological assessment, including and emphasizing the ripening time as the main parameter of grated Parmesan cheese. [ABSTRACT FROM AUTHOR]

Urea polyacrylamide gel electrophoresis combined with statistical methods was used to detect the addition of Ricotta to grated Parmigiano Reggiano cheese. The adulterated samples were prepared by adding grated Ricotta to grated Parmigiano Reggiano cheese in six different ratios. The logistic regression and rank correlation performed worse than the naïve Bayes classifier at identifying samples of grated Parmigiano Reggiano containing more than 20% Ricotta in terms of area under the receiver operating characteristic curve and classification accuracy. This work can be considered a pilot study to investigate the potential of machine‐learning methods to predict the authenticity of Parmigiano Reggiano cheese. [ABSTRACT FROM AUTHOR]

Fermentation is a promising solution to valorize cheese whey, the main by-product of the dairy industry. In Parmigiano Reggiano cheese production, natural whey starter (NWS), an undefined community of thermophilic lactic acid bacteria, is obtained from the previous day residual whey through incubation at gradually decreasing temperature after curd cooking. The aim of this study was to investigate the effect of fermentation regime (spontaneous (S) and NWS-inoculated (I-NWS)) on biofunctionalities and release of bioactive peptides during whey fermentation. In S and I-NWS trials proteolysis reached a peak after 24 h, which corresponded to the drop out in pH and the maximum increase in lactic acid. Biological activities increased as a function of fermentation time. NWS inoculum positively affected antioxidant activity, whilst S overcame I-NWS in angiotensin-converting enzyme (ACE) and DPP-IV (dipeptidyl peptidase IV) inhibitory activities. Peptidomics revealed more than 400 peptides, mainly derived from β-casein, κ-casein, and α-lactalbumin. Among them, 49 were bioactive and 21 were ACE-inhibitors. Semi-quantitative analysis strongly correlated ACE-inhibitory activity with the sum of the peptide abundance of ACE-inhibitory peptides. In both samples, lactotripeptide isoleucine-proline-proline (IPP) was higher than valine-proline-proline (VPP), with the highest content in S after 24 h of fermentation. In conclusion, we demonstrated the ability of whey endogenous microbiota and NWS to extensively hydrolyze whey proteins, promoting the release of bioactive peptides and improving protein digestibility.

Cheese is a worldwide produced and consumed commodity. There are many varieties of cheese from soft to hard, white to yellow, and fresh to aged after ripening. Especially, each category has its own producing technology. Many countries have labeled their most traditional cheese as Protective Designation of Origin (PDO). Moreover, several studies using advanced technologies, such as proteomics, have been performed to enhance labeling. In this review, broadly diffused and marketed, as well as Mediterranean countries' special interest in Mediterranean diet-related PDO cheeses have been chosen as a reference. The aim of this work was to highlight the use of proteomics methods to examine how cheese proteins and peptides rearrange after ripening and use of starters. Further, we aimed to examine what kind of proteins are produced. Finally, we focused on bioactive molecules in cheeses and distinction of the original product from its counterfeit. [ABSTRACT FROM AUTHOR]

Cheese microbiota contribute significantly to the final characteristics of cheeses due to the growth and interaction between cheese microorganisms during processing and ripening. For raw milk cheeses, such as Parmigiano Reggiano (PR), the microbiota derive from the raw milk itself, the dairy environment, and the starter. The process of cheese making and time of ripening shape this complex ecosystem through the selection of different species and biotypes that will drive the quality of the final product by performing functions of their metabolism such as proteolysis. The diversity in the final peptide and amino acid composition of the cheese is thus mostly linked to the diversity of this microbiota. The purpose of this study was to get more insight into the factors affecting PR cheese diversity and, more specifically, to evaluate whether the composition of the bacterial community of cheeses along with the specific peptide composition are more affected by the ripening times or by the cheese making process. To this end, the microbiota and the peptide fractions of 69 cheese samples (from curd to cheese ripened 24 months) were analyzed during 6 complete PR production cycles, which were performed in six different dairies located in the PR production area. The relation among microbial dynamics, peptide evolution, and ripening times were investigated in this unique and tightly controlled production and sampling set up. The study of microbial and peptide moieties in products from different dairies – from curd to at least 12 months, the earliest time from which the cheese can be sold, and up to a maximum of 24 months of ripening – highlighted the presence of differences between samples coming from different dairies, probably due to small differences in the cheese making process. Besides these differences, however, ripening time had by far the greatest impact on microbial dynamics and, consequently, on peptide composition.

Grana Padano (GP) and Parmigiano Reggiano (PR) are the two most important Italian PDO cheeses. To improve the environmental sustainability of their production, a Life Cycle Assessment (LCA) was completed on 84 dairy farms located in the province of Mantova (Northern Italy). In particular, 33 farms delivered milk for GP production, whereas 51 farms to dairies for PR production. In GP farms, maize silage represented 33.7% of total farmland and alfalfa represented 28.1%. While in PR farms, alfalfa represented 63.6% of total farmland. Fat and Protein Corrected Milk (FPCM) and Dairy Efficiency (DE, calculated as kg of produced FPCM per kg of DM intake) were different in the two production system: FPCM was 30.2 ± 4.32 kg/d in GP farms and 25.0 ± 4.71 kg/d in PR farms; DE was 1.35 ± 0.26 in GP farms, and 1.15 ± 0.22 in PR farms. Mitigation strategies to improve both environmental and economic sustainability were suggested focussing on forage crop production, milk production, herd management and off-farm purchased feed. From the preliminary results, there is evidence that improvements are needed. Climate Change (kg CO2 eq/kg FPCM) and Land Use (kg Carbon deficit/kg FPCM) were similar (1.38 ± 0.33 and 19.3 ± 7.08 for GP system; 1.46 ± 0.37 and 21.8 ± 11.4 for PR system). The most efficient farms in terms of milk production and DE generally showed the best environmental and economic sustainability, while the others show worse outcomes, mainly due to poor DE, livestock-management issues, feed purchase and ration composition.Highlights 84 farms producing milk for Grana Padano and Parmigiano Reggiano cheese were studied in the Province of Mantova. Life Cycle Assessment was used to quantify the environmental impacts of farms and statistical analysis was helpful to identify 6 clusters. Farm, animal and milk efficiencies were poor and mitigation strategies to improve the sustainability of milk production were suggested.

Cheese microbiota contribute significantly to the final characteristics of cheeses due to the growth and interaction between cheese microorganisms during processing and ripening. For raw milk cheeses, such as Parmigiano Reggiano (PR), the microbiota derive from the raw milk itself, the dairy environment, and the starter. The process of cheese making and time of ripening shape this complex ecosystem through the selection of different species and biotypes that will drive the quality of the final product by performing functions of their metabolism such as proteolysis. The diversity in the final peptide and amino acid composition of the cheese is thus mostly linked to the diversity of this microbiota. The purpose of this study was to get more insight into the factors affecting PR cheese diversity and, more specifically, to evaluate whether the composition of the bacterial community of cheeses along with the specific peptide composition are more affected by the ripening times or by the cheese making process. To this end, the microbiota and the peptide fractions of 69 cheese samples (from curd to cheese ripened 24 months) were analyzed during 6 complete PR production cycles, which were performed in six different dairies located in the PR production area. The relation among microbial dynamics, peptide evolution, and ripening times were investigated in this unique and tightly controlled production and sampling set up. The study of microbial and peptide moieties in products from different dairies – from curd to at least 12 months, the earliest time from which the cheese can be sold, and up to a maximum of 24 months of ripening – highlighted the presence of differences between samples coming from different dairies, probably due to small differences in the cheese making process. Besides these differences, however, ripening time had by far the greatest impact on microbial dynamics and, consequently, on peptide composition. [ABSTRACT FROM AUTHOR]

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Milk characteristics in terms of volatile compounds can influence the subsequent product characteristics and can give indications about metabolism. These features can strongly depend on feeding and management. In this perspective, the screening of milk samples intended for Parmigiano-Reggiano and Grana Padano located in Northern Italy was performed, focusing on a panel of volatile molecules. The work was carried out on a total number of 25 bovine milk samples for the production of Parmigiano-Reggiano and Grana Padano. Milk samples were collected from May to September and analyzed for volatile molecules using headspace gas chromatography mass spectrometry. A range of several volatile molecule classes was considered (aldehydes, ketones, alcohols, carboxylic acids, esters, aromatic hydrocarbons, solforates). Results showed a significant influence of the month and destination of milk due to the time period and subsequent use in cheesemaking (Parmigiano-Reggiano and Grana Padano). Significant differences between months were observed for all volatiles. These preliminary results indicate differences between the two types of milk due to the period and destination. The study of volatile molecules in milk will give important information about the physiology of milk and the evolution of dairy products. These features must be extended and confirmed by the sensory analysis of milk and derived products, leading to a more complete characterization of milk biology and derived products. [ABSTRACT FROM AUTHOR]

Raman spectroscopy, and handheld spectrometers in particular, are gaining increasing attention in food quality control as a fast, portable, non-destructive technique. Furthermore, this technology also allows for measuring the intact sample through the packaging and, with respect to near infrared spectroscopy, it is not affected by the water content of the samples. In this work, we evaluate the potential of the methodology to model, by multivariate data analysis, the authenticity of Parmigiano Reggiano cheese, which is one of the most well-known and appreciated hard cheeses worldwide, with protected denomination of origin (PDO). On the other hand, it is also highly subject to counterfeiting. In particular, it is critical to assess the authenticity of grated cheese, to which, under strictly specified conditions, the PDO is extended. To this aim, it would be highly valuable to develop an authenticity model based on a fast, non-destructive technique. In this work, we present preliminary results obtained by a handheld Raman spectrometer and class-modeling (Soft Independent Modeling of Class Analogy, SIMCA), which are extremely promising, showing sensitivity and specificity of 100% for the test set. Moreover, another salient issue, namely the percentage of rind in grated cheese, was addressed by developing a multivariate calibration model based on Raman spectra. It was possible to obtain a prediction error around 5%, with 18% being the maximum content allowed by the production protocol.

Parmigiano Reggiano cheese is one of the most appreciated Italian foods on account of its high nutrient content and taste. Due to its high cost, these characteristics make this product subject to counterfeiting in different forms. In this study, an approach based on an array of gas sensors has been employed to assess if it was possible to distinguish different samples based on their aroma. Samples were characterized in terms of rind percentage, seasoning, and rind working process. From the responses of the sensors, five features were extracted and the capability of these parameters to recognize target classes was tested with statistical analysis. Hence, the performance of the sensors’ array was quantified using artificial neural networks. To simplify the problem, a hierarchical approach has been used: three steps of classification were performed, and in each step one parameter of the grated cheese was identified (firstly, seasoning; secondly, rind working process; finally, rind percentage). The accuracies ranged from 88.24% to 100%.

Grana Padano (GP) and Parmigiano Reggiano (PR) are the two most important Italian PDO cheeses. To improve the environmental sustainability of their production, a Life Cycle Assessment (LCA) was completed on 84 dairy farms located in the province of Mantova (Northern Italy). In particular, 33 farms delivered milk for GP production, whereas 51 farms to dairies for PR production. In GP farms, maize silage represented 33.7% of total farmland and alfalfa represented 28.1%. While in PR farms, alfalfa represented 63.6% of total farmland. Fat and Protein Corrected Milk (FPCM) and Dairy Efficiency (DE, calculated as kg of produced FPCM per kg of DM intake) were different in the two production system: FPCM was 30.2 ± 4.32 kg/d in GP farms and 25.0 ± 4.71 kg/d in PR farms; DE was 1.35 ± 0.26 in GP farms, and 1.15 ± 0.22 in PR farms. Mitigation strategies to improve both environmental and economic sustainability were suggested focussing on forage crop production, milk production, herd management and off-farm purchased feed. From the preliminary results, there is evidence that improvements are needed. Climate Change (kg CO2 eq/kg FPCM) and Land Use (kg Carbon deficit/kg FPCM) were similar (1.38 ± 0.33 and 19.3 ± 7.08 for GP system; 1.46 ± 0.37 and 21.8 ± 11.4 for PR system). The most efficient farms in terms of milk production and DE generally showed the best environmental and economic sustainability, while the others show worse outcomes, mainly due to poor DE, livestock-management issues, feed purchase and ration composition. 84 farms producing milk for Grana Padano and Parmigiano Reggiano cheese were studied in the Province of Mantova. Life Cycle Assessment was used to quantify the environmental impacts of farms and statistical analysis was helpful to identify 6 clusters. Farm, animal and milk efficiencies were poor and mitigation strategies to improve the sustainability of milk production were suggested. [ABSTRACT FROM AUTHOR]

Parmigiano Reggiano is a typical Italian product known all over the world. Appreciated for its qualities, it is also one of the most counterfeit foods. To avoid this, two different techniques has been used: GC-MS with SPME and S3 gas sensor. With the first one, characteristic VOCs of different cheeses has been found. The most present classes are alcohols, aldehydes, hydrocarbons, esters and ketones, for a total of 58 compounds. Statistical analysis has shown that only 6 of them could be used to distinguish between Parmigiano Reggiano and its competitors. Instead, S3 is a new, rapid, economic and user-friendly approach. Assessing the variation of sensors resistances, it has been possible to discriminate between different kind of cheeses with an accuracy higher than 80%. Data analysis was performed considering PLS to have a visual representation of samples, the SFS algorithm for features selection and PLS-DA as classifier. [ABSTRACT FROM AUTHOR]

We investigated the effects of monensin controlled-release capsule (CRC; Kexxtone, Eli Lilly and Company Ltd., Indianapolis, IN) preventative ketosis treatment on the traditional cheesemaking process as well as the final characteristics of Parmigiano Reggiano (PR) cheese. The use of this prevention product to reduce the incidence of ketosis in transition dairy cows was approved by the European Medicines Agency in 2013. No previous studies are available concerning the effects of this treatment on prolonged-ripening cheese production such as PR. In PR cheese production, feed, feed additives, and cow treatments are strictly regulated to avoid any possible interference with traditional manufacturing processes. For these reasons, on 1 farm where all milk was used for PR cheese production, monensin CRC was administered to 33 cows, 21 d before calving in the monensin-treated group (TRT), whereas untreated cows with similar breed and parity characteristics constituted the control group (CTR). For 20 wk, milk obtained from each group and whey starter were separately managed and transported in the cheese factory, where 2 cheese wheels per group were produced daily, making 552 PR cheese wheels in total. Morning bulk tank milk composition, cheesemaking properties, and whey starter fermentation activities were analyzed twice a week. Every aspect of the cheesemaking process was recorded and the resulting cheese was evaluated after 36 h and 6, 12, and 18 mo from production for yield, texture defects, composition, and fatty acids profile. Milk from the 2 groups differed for somatic cell content (TRT = 3.04 vs. CTR = 4.06, somatic cell score), total bacterial count (TRT = 4.08 vs. CTR = 6.08 × 1,000 cfu/mL), titratable acidity (TRT = 3.66 vs. CTR = 3.72 Soxhlet-Henkel degrees/50 mL), and casein content percentage (TRT = 2.4 vs. CTR = 2.5%). Whey starter parameters were comparable between the 2 groups. Final cheese composition and organoleptic profile were not influenced by the treatment, except for C18:1 content being enhanced (TRT = 22.8 vs. CTR = 20.8% of fatty acids). Percentage of defected ripened cheese was significantly lower in the treated group, both at x-ray evaluation performed at 6 mo (TRT = 6.2 vs. CTR = 12.3%) and at the consortium inspection, performed at 12 mo of ripening (TRT = 1.5 vs. CTR = 6.5%). On the other hand, average cheese yield at 18 mo of ripening was partially reduced (TRT = 7.5 vs. CTR = 7.7%). Overall, the use of monensin CRC had no negative effect on the cheesemaking process, prolonged ripening cheese characteristics, milk composition, or whey starter quality. [ABSTRACT FROM AUTHOR]

Milk characteristics in terms of volatile compounds can influence the subsequent product characteristics and can give indications about metabolism. These features can strongly depend on feeding and management. In this perspective, the screening of milk samples intended for Parmigiano-Reggiano and Grana Padano located in Northern Italy was performed, focusing on a panel of volatile molecules. The work was carried out on a total number of 25 bovine milk samples for the production of Parmigiano-Reggiano and Grana Padano. Milk samples were collected from May to September and analyzed for volatile molecules using headspace gas chromatography mass spectrometry. A range of several volatile molecule classes was considered (aldehydes, ketones, alcohols, carboxylic acids, esters, aromatic hydrocarbons, solforates). Results showed a significant influence of the month and destination of milk due to the time period and subsequent use in cheesemaking (Parmigiano-Reggiano and Grana Padano). Significant differences between months were observed for all volatiles. These preliminary results indicate differences between the two types of milk due to the period and destination. The study of volatile molecules in milk will give important information about the physiology of milk and the evolution of dairy products. These features must be extended and confirmed by the sensory analysis of milk and derived products, leading to a more complete characterization of milk biology and derived products.

Parmigiano Reggiano (PR) is a raw-milk, hard cooked, long-ripened cheese of high quality and nutritional value. Long ripening times allow for extensive proteolysis of milk proteins to yield a number of peptides, some of which have potential healthy bioactive properties. This study aimed to: i) determine the peptide profile of PR cheese subjected to simulated gastrointestinal transit; ii) evaluate in vitro whether the peptides could support growth of beneficial microbial groups of the gut microbiota. PR samples were subjected to in vitro digestion, simulating oral, gastric, and duodenal transit. Liquid chromatography coupled with tandem mass spectrometry revealed that digestion caused the disappearance of the serum proteins and most of the original peptides, while 71 new peptides were found, all ranging from 2 to 24 residues. The digests were given as sole nitrogen source to pure cultures of Bifidobacterium (27 strains) and Lactobacillus (30 strains), and to bioreactor batch cultures of human gut microbiota. Most of bifidobacteria and lactobacilli grew more abundantly on PR digests than on the control peptone, and exhibited strain- or species-specific peptide preferences, as evidenced by principal component analysis. Bifidobacteria generally consumed a greater amount of peptides than lactobacilli, in terms of both the mean peptide consumption and the number of peptides consumed. For bifidobacteria, peptide preferences were very diverse, but a core of 10 peptides with 4 or 5 residues were consumed by all the strains. Lactobacilli behaved more homogenously and consumed nearly only the same 6 peptides, mostly dipeptides. The peptide preferences of the different groups of bifidobacteria and lactobacilli could not be ascribed to features such as the length of the peptide or the abundance of residues with peculiar properties (hydrophobicity, polarity, charge) and likely depend on specific proteases and/or peptide transporters preferentially recognizing specific sequence motifs. The cultures of human colonic microbiota confirmed that PR digest promoted the growth of commensal bifidobacteria. This study demonstrated that peptides derived from simulated gastrointestinal digestion of PR supported the growth of most lactobacilli and bifidobacteria. [ABSTRACT FROM AUTHOR]

Os queijos de denominação geral de Grana (queijos duros), na Itália, apresentam renome mundial: o Grana Padano e o Parmigiano Reggiano. Ambos são elaborados com leite de vaca e possuem grande semelhança ente si, mas não se trata do mesmo queijo. O objetivo desse trabalho é apresentar uma revisão sobre os queijos Grana Italianos.

This study investigated aflatoxin B1 (AFB1) contamination in dairy cow feed and the risk management of AFB1 content in concentrates undertaken by feed industries in the Parmigiano Reggiano area. Data on aflatoxin contamination risk management applied in 29 feed industries were collected and the AFB1 content of 70 feed samples was analysed. Data were collected within the framework of a quality control programme promoted by the Parmigiano Reggiano Consortium in 2013 and 2014. Audit results showed that the control procedures to prevent AFB1 contamination mainly focused on maize and its by-products. AFB1 concentration resulted lower than 5 ppb [legal European Union (EU) limit] in all samples; in one out of 70 samples, AFB1 content was 3.8 ppb and in all the other samples it was lower than 3 ppb. Results showed that AFB1 risk management applied by Italian feed industries effectively monitors AFB1 levels in feed below the EU legal limit.

A novel nanowire-based sensor device is presented for the quality evaluation of the Italian Parmigiano Reggiano (PR) cheese. As a Protected Designation of Origin (PDO) trademark product, the cheese should follow stringent manufacturing guidelines and must exhibit the representative aromatic volatile organic compounds (VOCs) biomarkers and sensory profile. Any alteration in these features indicates an adulteration of the product as many low quality and substandard PR cheeses are widely available in the market. Different types of grated cheese samples were analyzed and the VOCs were characterized by evaluating the samples with the portable sensor device S3 and Gas Chromatography–Mass Spectrometry technique. The ability of S3 device to distinguish the cheese quality demonstrates its suitability for monitoring the quality control and authenticity of PR cheese. [ABSTRACT FROM AUTHOR]

To determine the originality of a typical Italian Parmigiano Reggiano cheese, it is crucial to define and characterize its quality, ripening period, and geographical origin. Different analytical techniques have been applied aimed at studying the organoleptic and characteristic volatile organic compounds (VOCs) profile of this cheese. However, most of the classical methods are time consuming and costly. The aim of this work was to illustrate a new simple, portable, fast, reliable, non-destructive, and economic sensor device S3 based on an array of six metal oxide semiconductor nanowire gas sensors to assess and discriminate the quality ranking of grated Parmigiano Reggiano cheese samples and to identify the VOC biomarkers using a headspace SPME-GC-MS. The device could clearly differentiate cheese samples varying in quality and ripening time when the results were analyzed by multivariate statistical analysis involving principal component analysis (PCA). Similarly, the volatile constituents of Parmigiano Reggiano identified were consistent with the compounds intimated in the literature. The obtained results show the applicability of an S3 device combined with SPME-GC-MS and sensory evaluation for a fast and high-sensitivity analysis of VOCs in Parmigiano Reggiano cheese and for the quality control of this class of cheese. [ABSTRACT FROM AUTHOR]

Parmigiano Reggiano (PR) cheese is one of the most important Italian Protected Designation of Origin (PDO) cheeses and it is exported worldwide. As a PDO, the product is supposed to have distinctive sensory characteristics. In this work we present the use of the Small Gas Sensor System (S3) device for the identification of specific PR markers, as compared to classical chemical techniques, such as Gas chromatography-mass spectrometry solid-phase microextraction (SPME-GC-MS). Markers are used to determine the percent of grated pulp and rind commercially utilized. The S3 device is equipped with an array of six metal oxide semiconductor (MOX) gas sensors, three of them with a nanowire (NW) morphology and the other three in the form of thin films. PDO can cover grated PR cheese as well, but only if made with whole cheese. Grated PR cheese must be characterized by the absence of additives and no more than 18% crust. The achieved results strongly encourage the use of S3 for a rapid identification of the percentage of grated PR. [ABSTRACT FROM AUTHOR]

Thirty-nine strains of Kluyveromyces marxianus from Pecorino di Farindola cheese in comparison with 3 strains from Parmigiano Reggiano cheese, 1 from fermented milk, 3 from cow whey and two type strains K. marxianus CBS 834 T and Kluyveromyces lactis CBS 683 T were tested for genetic and metabolic characteristics. Intraspecific diversity of chromosome arrangements was evaluated by pulsed field gel electrophoresis (PFGE) analysis. Among K. marxianus strains chromosome polymorphisms were evident with 11 patterns that differed in size and number of the chromosomal bands. The number of the bands varied from 4 to 7 with sizes ranging from about 1.0 to 2.7 Mb. Twelve strains were selected for determining their growth capacity and volatile compound production in two wheys (raw cheese whey and ricotta cheese whey) under limited oxygen availability. The growth kinetics highlighted four different biotypes and the influence of whey composition on K. marxianus development. The main volatile compounds detected after the growth were alcohols, acids, esters, ketones and aldehydes. Ethanol was the most abundant in both wheys. Aldehydes and other minor compounds were produced only when the strains were inoculated in ricotta cheese whey, while esters, butanoic, decanoic and octanoic acids were qualitatively and quantitatively more present in raw cheese whey. This study highlights a great genetic and metabolic biodiversity within Pecorino di Farindola K. marxianus strains and it could be exploited to improve the knowledge of this yeast for biotechnological uses. [ABSTRACT FROM AUTHOR]

The occurrence of 8 bovine casein-derived peptides (VPP, IPP, RYLGY, RYLG, AYFYPEL, AYFYPE, LHLPLP, and HLPLP) reported as angiotensin converting enzyme-inhibitors (ACE-I) was investigated in the 3-kDa ultrafiltered water-soluble extract (WSE) of Parmigiano Reggiano (PR) cheese samples by ultraperformance liquid chromatography coupled to highresolution mass spectrometry via an electrospray ionization source. Only VPP, IPP, LHLPLP, and HLPLP were revealed in the WSE, and their total amount was in the range of 8.46 to 21.55 mg/kg of cheese. Following in vitro static gastrointestinal digestion, the same ACEI peptides along with the newly formed AYFYPEL and AYFYPE were found in the 3 kDa WSE of PR digestates. Digestates presented high amounts (1,880-3,053 mg/kg) of LHLPLP, whereas the remaining peptides accounted for 69.24 to 82.82 mg/kg. The half-maximal inhibitory concentration (IC50) values decreased from 7.92 ± 2.08 in undigested cheese to 3.20 ± 1.69 after in vitro gastrointestinal digestion. The 3-kDa WSE of digested cheeses were used to study the transport of the 8 ACE-I peptides across the monolayers of the Caco-2 cell culture grown on a semipermeable membrane of the transwells. After 1 h of incubation, 649.20 ± 148.85 mg/kg of LHLPLP remained in the apical compartment, whereas VPP, IPP, AYFYPEL, AYFYPE, and HLPLP accounted in total for less than 36.78 mg/kg. On average, 0.6% of LHLPLP initially present in the digestates added to the apical compartment were transported intact to the basolateral chamber after the same incubation time. Higher transport rate (2.9%) was ascertained for the peptide HLPLP. No other intact ACE-I peptides were revealed in the basolateral compartment. For the first time, these results demonstrated that the ACE-I peptides HLPLP and LHLPLP present in the in vitro digestates of PR cheese are partially absorbed through an in vitro model of human intestinal epithelium. [ABSTRACT FROM AUTHOR]

Biogas production is increasing as a sustainable energy supply, with digestate resulting as a by-product of biogas plants. As a result, the high concentration of Clostridium spp. in digestate became a concern in dairy farming areas. Clostridium spores can contaminate soils and crops when digestate is used as fertilizer, causing a conflictual cohabitation of biogas with traditional cheese productions. In order to solve the problem, this study aimed to search for a technical solution enabling either a drastic reduction or the elimination of the content of Clostridium spp. within digestate. Results showed a complete elimination of Clostridium spp. in pelleted stored solid digestate; in addition, pelleting caused a reduction of pH and water mass fraction in terms of fresh weight, and a concentration of mineral nutrients compared to stored solid digestate. Pellet can represent a possible sustainable solution both in reducing potential risks linked to the presence of Clostridium spp. in digestate and in improving the transportation and distribution of high-value fertilizer. Hence, pelleting of solid digestate could offer a simple and efficient method to allow cohesistence of biogas plants and dairy farming. [ABSTRACT FROM AUTHOR]

To determine the originality of a typical Italian Parmigiano Reggiano cheese, it is crucial to define and characterize its quality, ripening period, and geographical origin. Different analytical techniques have been applied aimed at studying the organoleptic and characteristic volatile organic compounds (VOCs) profile of this cheese. However, most of the classical methods are time consuming and costly. The aim of this work was to illustrate a new simple, portable, fast, reliable, non-destructive, and economic sensor device S3 based on an array of six metal oxide semiconductor nanowire gas sensors to assess and discriminate the quality ranking of grated Parmigiano Reggiano cheese samples and to identify the VOC biomarkers using a headspace SPME-GC-MS. The device could clearly differentiate cheese samples varying in quality and ripening time when the results were analyzed by multivariate statistical analysis involving principal component analysis (PCA). Similarly, the volatile constituents of Parmigiano Reggiano identified were consistent with the compounds intimated in the literature. The obtained results show the applicability of an S3 device combined with SPME-GC-MS and sensory evaluation for a fast and high-sensitivity analysis of VOCs in Parmigiano Reggiano cheese and for the quality control of this class of cheese.

New Zealand is a leader in the global dairy industry. Milk powder is the principal export product, but there is also a prominent cheese manufacturing industry, catering more for the domestic market. The Selected Ion Flow Tube-Mass Spectrometric (SIFT-MS) technique was used to compare 4 New Zealand cheeses marketed as 'parmesan' with 4 Italian Parmigiano Reggiano and Grana Padano cheeses. The cheese headspace was analyzed in real time without any sample preconcentration. Total of 38 volatile compounds in the cheese headspace were monitored with headspace concentrations varying between single digit parts per billion (ppb) to tens of parts per million (ppm). When the results were subjected to multivariate statistical analysis, a clear discrimination was found between the New Zealand 'parmesan' and Italian cheeses based solely on the measured concentrations of these volatile compounds. If the volatile compounds used in the analyses were restricted to known odor-active compounds in Parmigiano Reggiano cheese, the ability to discriminate between the cheeses was maintained. The analyses also showed that it was possible to clearly differentiate between the different processing plants in individual countries. Important discriminatory volatiles in the samples tested were butanoic acid and phenylacetaldehyde for discriminating between Italian cheeses and ethyl butyrate, acetaldehyde and methylbutanals between New Zealand cheeses. We conclude that the New Zealand 'parmesans' do not provide a good representation of the aroma of Italian 'parmesans.' Practical Application: SIFT-MS has been shown to clearly differentiate both country of origin and the manufacturer of 'parmesan' cheeses made in Italy and New Zealand based on differences in volatile organic compounds. Thus this method will have benefit for use in the quality control of 'parmesan' and other cheese varieties. [ABSTRACT FROM AUTHOR]

Abstract: Herein, the potential of Fourier transform infrared spectroscopy (FTIR) spectroscopy coupled with principal component analysis and artificial neural networks to discriminate butters obtained from milk creams collected in different seasons (spring, summer or winter) and produced from two feeding regimens (traditional or unifeed) in the Parmigiano Reggiano cheese area was investigated. The predictive ability of neural networks to predict the season of production was 100%. The favorable results obtained by artificial neural network (ANN) analysis were in agreement with those reported by principal component analysis (PCA) analysis. The ability of ANNs to predict the feeding regimens was 90.0%, 75.0% and 75.0%, respectively, for samples collected in spring, summer and winter. These results also confirm that the method is highly suitable for its intended purpose. [Copyright &y& Elsevier]

Abstract: In compliance with the European law (EC No. 510/2006), geographical indications and designations of origin for agricultural products and foodstuffs must be protected against mislabelling. This is particularly important for PDO hard cheeses, as Parmigiano Reggiano, that can cost up to the double of the no-PDO competitors. This paper presents two statistical models, based on isotopic and elemental composition, able to trace the origin of cheese also in grated and shredded forms, for which it is not possible to check the logo fire-marked on the rind. One model is able to predict the origin of seven types of European hard cheeses (in a validation step, 236 samples out of 240 are correctly recognised) and the other specifically to discriminate the PDO Parmigiano Reggiano cheese from 9 European and 2 extra-European imitators (260 out of 264 correct classifications). Both models are based on Random Forests. The most significant variables for cheese traceability common in both models are δ 13C, δ 2H, δ 15N, δ 34S and Sr, Cu, Mo, Re, Na, U, Bi, Ni, Fe, Mn, Ga, Se, and Li. These variables are linked not only to geography, but also to cow diet and cheese making processes. [Copyright &y& Elsevier]

Osteoporosis is a multifactorial disease characterized by loss of bone mass and microarchitectural deterioration of bone tissue, which leads to a consequent increase in the risk of skeletal fractures. Diet awakes a critical interest in osteoporosis, because it is one of the few determinants that can be safely modified. A healthy well balanced nutrition can play an important role in prevention and pathogenesis of osteoporosis, but also in support of a pharmacological therapy. Numerous evidences have already established that dietary calcium, proteins and vitamin D are essential nutrients for achieved peak bone mass and maintaining skeletal health. Dairy products, by providing both calcium and proteins, represent the optimal source of highly bioavailable nutrients for bone health. Among dairy foods in particular cheese results one of the major source of calcium in the adults western diet and also in the Italian adults diet. Parmigiano Reggiano cheese is an homemade Italian food whose denomination "Protected Designation of Origin" is linked to an artisanal manufacturing process in limited geographic area of Northern Italy and is an optimal source of essential nutrients for acquisition and maintenance of bone health. Parmigiano Reggiano is a cheese easy digested, for the presence of ready to use proteins and lipids, lactose free, rich in calcium, with possible prebiotic and probiotic effect. On the basis of its nutritional characteristics and of its easy digestibility Parmigiano Reggiano cheese is recommended in all feeding age groups. [ABSTRACT FROM AUTHOR]

Aims: The aim of this work was to obtain a deeper insight into the knowledge of microbial composition of Parmigiano Reggiano natural whey starters through different culture-independent methods. Methods and Results: Eighteen different Parmigiano Reggiano natural whey starters sampled from three different provinces of this cheese production area and the nonacidified wheys from which they arose have been studied by length heterogeneity polymerase chain reaction (LH-PCR) and fluorescent in situ hybridization (FISH). A high microbial composition variability between different samples has been observed. Conclusions: Revealing different images of the same community, LH-PCR and FISH have given a more accurate view of the not well-known Parmigiano Reggiano whey starter ecosystem. Significance and Impact of the Study: New lights have been shed on Parmigiano Reggiano natural whey starters microbial composition, highlighting how culture-independent approach could be used and improved to study this and other food ecosystems. [ABSTRACT FROM AUTHOR]

Abstract: The diversity of dominant lactic acid bacteria population in 12 months ripened Parmigiano Reggiano cheeses was investigated by a polyphasic approach including culture-dependent and independent methods. Traditional plating, isolation of LAB and identification by 16S rDNA analysis showed that strains belonging to Lactobacillus casei group were the most frequently isolated. Lactobacillus helveticus, Lactobacillus delbrueckii subsp. lactis, Lactobacillus parabuchneri, and Lactobacillus buchneri species were detected with lower frequency. PCR-denaturing gradient gel electrophoresis (DGGE) applied to DNA extracted directly from cheese samples and sequencing of rDNA amplicons confirmed the complex microbiological pattern of LAB in ripened Parmigiano Reggiano cheeses, with the significant exception of the Lactobacillus fermentum species, which dominated in several samples, but was not detected by cultivation. The present combination of different approaches can effectively describe the lactic acid bacteria population of Parmigiano Reggiano cheese in advanced stages of ripening, giving useful information for elucidating the role of LAB in determining the final cheese quality. [Copyright &y& Elsevier]

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Abstract: Samples of Italian Parmigiano Reggiano cheese of different ripening stages (14, 24 and 30 months) were analyzed by 1H NMR spectroscopy and the water-soluble metabolites content compared with samples of “Grana type” cheese from east Europe countries. Different multivariate statistical protocols were examined to build up a stable model for both ripening and geographical discrimination among the samples. The discriminant approach revealed a larger metabolite content increasing with ripening process; in particular younger samples were characterized by a higher content in leucine and isoleucine, while 30 months samples by a larger extent of threonine. The use of a classification approach resulted in a better discrimination of geographical samples. Foreign “Grana type” samples resulted well differentiated with respect to all Italian samples of Parmigiano Reggiano cheese. Foreign samples were characterized by a large amount of leucine and isoleucine, compounds that typified young Italian samples (14 months), and also by lactate, butanoate and acetate, thus suggesting short ripening for foreign samples. Parmigiano Reggiano samples were instead characterized by a large amount of all other compounds, in particular by threonine, which typified old samples (30 months), and other amino acids. [Copyright &y& Elsevier]

Abstract: Traceability is becoming a method of providing safer food supplies and of connecting producers and consumers. Recent diseases such as bovine spongiform encephalitis (BSE) and the questions concerning genetically modified organism (GMO) mean systems that enable control of each link in the food chain have become particularly relevant. Furthermore, although EU law no. 178 came into effect on the 1st January 2005, at the time of writing the regulatory situation is very confused. The aim of this paper is to analyze legal and regulatory aspects of food traceability, and to provide a general framework for the identification of fundamental mainstays and functionalities in an effective traceability system. Possible technical resources were clarified by analyzing assessment criteria obtained from studies of alphanumerical codes, bar codes, and radio frequency identification (RFID). Finally, the paper presents the traceability system used by Parmigiano Reggiano (the famous Italian cheese) which was developed using the proposed general framework. Based on an integration of alphanumerical codes and RFID technology, the system is working well with very good results for both cheese producers and consumers. Some interesting observations concerning development trends and traceability system costs close the paper. [Copyright &y& Elsevier]

Abstract: This study aimed to compare phenotypic and genetic characteristics of Lactobacillus rhamnosus strains isolated at the end of the ripening of Parmigiano Reggiano cheese and to investigate an important prerequisite of probiotic interest, such as the capability to survive at low pH and in presence of bile salts. The use of API 50 CH, RAPD-PCR analysis and species-specific PCR allowed to ascertain the identity of 63 L. rhamnosus strains. Three L. rhamnosus strains isolated from Parmigiano Reggiano cheese, L. rhamnosus ATCC 7469T and the commercial strain L. GG were assayed to estimate the resistance to various stress factors reproducing in vitro some conditions of the gastro-intestinal environment such as low pH and different amounts of bile salts and acids. The behaviour of almost all the tested strains isolated from Parmigiano Reggiano cheese resulted analogous to that showed by L. GG. [Copyright &y& Elsevier]

Abstract Aims: Lactobacillus helveticus is the dominant microflora of the natural whey starters used for Parmigiano Reggiano cheese making. The aim of this work was to study the biodiversity of different strains of Lact. helveticus present in six cultures and to compare them with strains of the same species previously isolated from natural whey cultures used for Grana Padano and Provolone cheeses. Methods and Results: Twenty different biotypes of Lact. helveticus strains were identified combining the results deriving from SDS-PAGE of cell surface proteins and PCR fingerprinting using M13 as a primer. The biotypes were present in varying amounts in the six natural whey starters and the biodiversity was demonstrated not only within the whey cultures, but also between the whey cultures. Conclusions: Lact. helveticus strains isolated from Parmigiano Reggiano whey cultures analysed by PCR M13, SDS-PAGE and RFLP were distinguishable from Lact. helveticus strains of different dairy origin, namely Grana Padano and Provolone natural whey starters. Significance and Impact of the Study: The presence of different Lact. helveticus biotypes seems to be related to the specific ecosystem of cheesemaking and may be considered as one of the elements contributing to the typicality of Parmigiano Reggiano cheese. [ABSTRACT FROM AUTHOR]

This study investigated aflatoxin B1 (AFB1) contamination in dairy cow feed and the risk management of AFB1 content in concentrates undertaken by feed industries in the Parmigiano Reggiano area. Data on aflatoxin contamination risk management applied in 29 feed industries were collected and the AFB1 content of 70 feed samples was analysed. Data were collected within the framework of a quality control programme promoted by the Parmigiano Reggiano Consortium in 2013 and 2014. Audit results showed that the control procedures to prevent AFB1 contamination mainly focused on maize and its by-products. AFB1 concentration resulted lower than 5 ppb [legal European Union (EU) limit] in all samples; in one out of 70 samples, AFB1 content was 3.8 ppb and in all the other samples it was lower than 3 ppb. Results showed that AFB1 risk management applied by Italian feed industries effectively monitors AFB1 levels in feed below the EU legal limit. [ABSTRACT FROM AUTHOR]