Ingested versus inhaled limonene in sheep: A pilot study to explore potential different transfer to the mammary gland and effects on milk and Caciotta cheese aroma.

Concentration is a key determinant in the overall positive impact of terpenes on milk and cheese aroma; additionally, route of intake may affect the achievable concentrations of dietary terpenes in milk and cheese. In this study, we explored the possibility that the amount of the monoterpene limonene transferred to sheep milk and its corresponding cheese could differ depending on the route of intake and that the aroma profile of these products could also differ. To this aim, 12 lactating dairy ewes were repeatedly exposed to limonene by the oral or respiratory route during a 48-h test period, according to a 3 × 3 Latin square experimental design. Limonene content was measured in individual and bulk milk samples, in 1-d-old and 15-d-old Caciotta cheese obtained from that milk, in the related whey and curd, and in the air inhaled by the ewes in the respiratory treatment group (to obtain an estimate of the dose actually supplied by this route). Bulk milk and fresh (1-d-old) cheese underwent sensory analysis by ortho-olfactory evaluation. Both intake routes demonstrated transfer of limonene to milk, but the respiratory route transferred limonene with greater efficiency than the oral route. Moreover, according to the protocol used in this study, a short period of respiratory exposure induced a slightly higher limonene content in milk compared with oral exposure. As to the fate of limonene during cheesemaking, an important part of it was lost into the whey, perhaps through volatilization. The differences between milk and cheese tended to dissipate in curd and fresh cheese and disappeared completely after 15 d of ripening. Finally, it was possible to distinguish between the 2 routes of limonene intake using sensory analysis, even though no direct relationship was identified between the different aroma profiles of milks and cheeses from the oral and respiratory groups and their respective limonene contents. Overall, our results expand current knowledge on the biological pathways of terpene transfer from feed to sheep milk and cheese, as well as on the role played by terpenes in the formation of aroma in these products. Our observations may contribute to future development of strategies for external control and better standardization of the presence of odor compounds in milk and cheese from dairy ruminants.

Short communication: Sensory characteristics and volatile organic compound profile of high-moisture mozzarella made by traditional and direct acidification technology.

In the present study, the sensory characteristics and the volatile organic compound (VOC) profiles of high-moisture mozzarella made by different acidification techniques were compared. The cheeses were manufactured at the same dairy by fermentation by autochthonous natural whey starter (traditional backslopping method) and direct acidification with citric acid (the most used industrial technology). Three cheesemaking trials were performed from February to June using raw milk from a single farm. The mozzarella samples were subjected to assessment of the chemical, microbiological, and sensory characteristics and to VOC analysis by headspace solid-phase microextraction coupled to gas chromatography-mass spectrometry. The relevant microbiological differences found between the 2 types of products were reflected in different sensory and VOC profiles. The cheeses were clearly discriminated by the panel, and traditional mozzarella had higher intensity attributes. The most discriminating descriptors were elasticity, overall odor and taste intensity, sour milk and fruity/vegetable odors, sour taste, and aftertaste. Even though some variability was observed among the trials, the VOC profile of traditional product was always much more complex than that obtained by direct acidification. In both products the carbonyl compounds were the most abundant chemical class, accounting for about 50% of the total. In detail, 51 compounds were identified in the entire set of samples, and their contribution to cheese aroma was roughly estimated by calculating the odor activity values on the basis of the odor thresholds available in the literature. The results allowed hypothesizing that only 12 of them could play a primary role. The most important among the odor-active compounds was 3-methyl-butanal that can both derive from metabolism of lactic acid bacteria and yeasts. The results of the study may be very useful in view of European Union PDO labeling of the traditional product, in terms of protecting it from imitations.

Production of cheese from donkey milk as influenced by addition of transglutaminase.

Donkey milk is characterized by low contents of total solids, fat, and caseins, especially κ-casein, which results in formation of a very weak gel upon renneting. The objective of this study was to evaluate the effect of fortification of donkey milk with microbial transglutaminase (MTGase) for cheesemaking in relation to different enzyme addition protocols (patterns, PAT). Four independent trials were performed using MTGase (5.0 U/g of milk protein) according to the following experimental patterns: control (no MTGase addition); MTGase addition (40°C) 15 min before starter inoculation (PAT1); addition of MTGase to milk simultaneously with starter culture (40°C) (PAT2); and MTGase addition simultaneously with rennet (42°C) in acidified milk (pH 6.3) (PAT3). Evolution of pH during acidification, cheesemaking parameters, and proximal composition and color of cheese at 24 h were recorded. The protein fractions of cheese and whey were investigated by urea-PAGE and sodium dodecyl sulfate-PAGE. Addition of MTGase had no significant effect on moisture, protein, fat, or cheese yield. The addition of MTGase with rennet (PAT3) improved curd firmness compared with the control. Among the different patterns of MTGase addition, PAT3 reduced gel formation time, time between rennet addition and cheese molding, and weight loss of cheese at 24 h. The PAT3 treatment also resulted in the lowest lightness and highest yellowness color values of the cheese. Sodium dodecyl sulfate-PAGE of cheeses revealed that MTGase modified the protein pattern in the high-molecular-weight zone (range 37-75 kDa) compared with the control. Of the MTGase protocols, PAT3 showed better casein retention in cheese, as confirmed by the lanes of α- and ß-caseins in the electropherogram of the whey, which was subtler for this protocol. In conclusion, MTGase may be used in cheese production from donkey milk to improve curd firmness; MTGase should be added simultaneously with the rennet.

Short communication: Chemical-sensory and volatile compound characterization of ricotta forte, a traditional fermented whey cheese.

Ricotta forte is a traditional whey cheese, obtained through natural fermentation of fresh ricotta, that is getting increasing attention by food traders. In view of possible initiatives for its valorization, the chemical and sensory characteristics were investigated. Samples were obtained from 14 different manufacturer, and were subjected to chemical, biochemical, volatile organic compound, and sensory analyses. All samples presented low pH with high moisture (62-66%) and fat content (57-60% on dry matter). From a biochemical point of view, the electrophoretic patterns evidenced that ß-lactoglobulin was the main protein present at all sample ages. Only intermediate levels of proteolysis (20.69% ripening index) took place during aging, whereas the main biochemical event in this dairy product was lipolysis (2.10 mEq/g of acid degree value). Accordingly, free fatty acids dominated the volatile organic compound profile and strongly influenced the sensory characteristics with flavor described as rancid, pungent, acrid, and smelly feet: all associated with short-chain fatty acids such as acetic, propionic, butyric, and caproic. Finally, the sample age did not influence chemical composition, whereas it had significant effect on lipolysis and flavor intensity.

Comparison of HPLC-RI, LC/MS-MS and enzymatic assays for the analysis of residual lactose in lactose-free milk.

Lactose intolerance is the decreased ability to digest lactose, and the population involved is rapidly increasing all over the world. Different procedures have been reported in the literature to quantify lactose in dairy products, but the official method of analysis is based on enzymatic assay. In this paper, the effectiveness of two enzymatic kits in detecting residual lactose in lactose-free milk was investigated, and a comparison with two alternative chromatographic methods was done. The investigation used several samples of UHT milk containing different levels of lactose, and the results highlighted the inadequacy of the enzymatic assays and of the HPLC-RI method to analyse lactose-free milk. An LC-MS/MS method using the formate adduct was developed, and it allowed quantitation of lactose and lactulose in all samples at a high level of precision and repeatability.

Short communication: Chemical and sensory characteristics of Canestrato di Moliterno cheese manufactured in spring.

Canestrato di Moliterno is an Italian Protected Geographical Indication hard cheese, made in winter and spring from a mixture of ewe and goat milks, that has been poorly investigated. The present study was aimed at characterizing the cheese made in the warm season. Two series of samples, ripened in traditional rooms called fondaco as indicated in the official protocol of production, were taken from the main certified producers. The cheeses were analyzed for gross composition; proteolysis and lipolysis; volatile fraction; and organoleptic features. Gross composition was not completely homogeneous among the samples, but primary proteolysis and lipolysis were quite uniform. We observed variations in secondary proteolysis, likely caused by fluctuations in environmental conditions in the fondaco. The sensory profiles of the samples were homogeneous: the cheese was soluble, greasy, and adhesive, with a sheepfold and buttery odor. The main taste attributes were fermented, pungent, and bitter. Overall, the results of this study provide an initial contribution to the characterization of Canestrato di Moliterno, and could be used to improve marketing strategies.

Production technology and characterization of Fior di latte cheeses made from sheep and goat milks.

Innovation in the small ruminant dairy sector faces structural challenges because dairies are often involved in breeding and produce cheeses that appeal essentially to local markets using traditional technologies and facilities. An investigation was carried out to produce Fior di latte, a traditional, soft pasta filata cheese, from sheep and goat milks at the farm level. Fior di latte is an Italian high-moisture, round mozzarella currently produced from cow and water buffalo milks; it is very popular in Europe. Cheesemaking trials were performed and the most appropriate technology proved to be a combination of direct acidification and lactic fermentation, with some modifications to the milk coagulation phase. The gross composition of the experimental cheeses was similar to that of bovine Fior di latte, and the overall hygienic quality was satisfactory even though the milk had not been pasteurized. The new cheeses were similar in appearance to the bovine type, but some specific features were detected. Besides the typical "goaty" and "sheepy" flavors, some novel and distinctive descriptors of odor, flavor, and texture were noted. Our experiment showed that good quality Fior di latte cheese that complies with microbiological requirements of the European legislation can be obtained from sheep and goat milks by appropriately modifying the cheesemaking technology.

Manufacture of Fior di Latte cheese by incorporation of probiotic lactobacilli.

This work aimed to select heat-resistant probiotic lactobacilli to be added to Fior di Latte (high-moisture cow milk Mozzarella) cheese. First, 18 probiotic strains belonging to Lactobacillus casei, Lactobacillus delbrueckii ssp. bulgaricus, Lactobacillus paracasei, Lactobacillus plantarum, Lactobacillus rhamnosus, and Lactobacillus reuteri were screened. Resistance to heating (65 or 55°C for 10 min) varied markedly between strains. Adaptation at 42°C for 10 min increased the heat resistance at 55°C for 10 min of all probiotic lactobacilli. Heat-adapted L. delbrueckii ssp. bulgaricus SP5 (decimal reduction time at 55°C of 227.4 min) and L. paracasei BGP1 (decimal reduction time at 55°C of 40.8 min) showed the highest survival under heat conditions that mimicked the stretching of the curd and were used for the manufacture of Fior di Latte cheese. Two technology options were chosen: chemical (addition of lactic acid to milk) or biological (Streptococcus thermophilus as starter culture) acidification with or without addition of probiotics. As determined by random amplified polymorphic DNA-PCR and 16S rRNA gene analyses, the cell density of L. delbrueckii ssp. bulgaricus SP5 and L. paracasei BGP1 in chemically or biologically acidified Fior di Latte cheese was approximately 8.0 log(10)cfu/g. Microbiological, compositional, biochemical, and sensory analyses (panel test by 30 untrained judges) showed that the use of L. delbrueckii ssp. bulgaricus SP5 and L. paracasei BGP1 enhanced flavor formation and shelf-life of Fior di Latte cheeses.

Biochemical patterns in ovine cheese: influence of probiotic strains.

This study was undertaken to evaluate the effect of lamb rennet paste containing probiotic strains on proteolysis, lipolysis, and glycolysis of ovine cheese manufactured with starter cultures. Cheeses included control cheese made with rennet paste, cheese made with rennet paste containing Lactobacillus acidophilus culture (LA-5), and cheese made with rennet paste containing a mix of Bifidobacterium lactis (BB-12) and Bifidobacterium longum (BB-46). Cheeses were sampled at 1, 7, 15, and 30 d of ripening. Starter cultures coupled with probiotics strains contained in rennet paste affected the acidification and coagulation phases leading to the lowest pH in curd and cheese containing probiotics during ripening. As consequence, maturing cheese profiles were different among cheese treatments. Cheeses produced using rennet paste containing probiotics displayed higher percentages of alpha(S1)-I-casein fraction than traditional cheese up to 15 d of ripening. This result could be an outcome of the greater hydrolysis of alpha-casein fraction, attributed to higher activity of the residual chymosin. Further evidence for this trend is available in chromatograms of water-soluble nitrogen fractions, which indicated a more complex profile in cheeses made using lamb paste containing probiotics versus traditional cheese. Differences can be observed for the peaks eluted in the highly hydrophobic zone being higher in cheeses containing probiotics. The proteolytic activity of probiotic bacteria led to increased accumulation of free amino acids. Their concentrations in cheese made with rennet paste containing Lb. acidophilus culture and cheese made with rennet paste containing a mix of B. lactis and B. longum were approximately 2.5 and 3.0 times higher, respectively, than in traditional cheese. Principal component analysis showed a more intense lipolysis in terms of both free fatty acids and conjugated linoleic acid content in probiotic cheeses; in particular, the lipolytic pattern of cheeses containing Lb. acidophilus is distinguished from the other cheeses on the basis of highest content of health-promoting molecules. The metabolic activity of the cheese microflora was also monitored by measuring acetic, lactic, and citric acids during cheese ripening. Cheese acceptability was expressed for color, smell, taste, and texture perceived during cheese consumption. Use of probiotics in trial cheeses did not adversely affect preference or acceptability; in fact, panelists scored probiotic cheeses higher in preference over traditional cheese, albeit not significantly.

Effects of olive maturation and stoning on quality indices and antioxidant content of extra virgin oils (cv. Coratina) during storage.

UNLABELLED: Quality indices, antioxidant compounds, and antioxidant activities of extra-virgin oils from Coratina olives were evaluated during a 12-mo storage. Whole and stoned olives, picked at 2 different maturation index (MI), were submitted to malaxation for 45 min and extracted by a 3-phase continuous system. A 90-min malaxation trial was also performed for the stoned olives. The following parameters were monitored: free acidity, peroxide value, K(232) and K(270) indices, sensory profile, total phenolic content (TPC), phenolic profiles, tocopherol compounds, and antioxidant activity (AA). The highest TPC, AA, and sensory score were found for the oils obtained by olives picked at low MI and by stoned olives. After 12 mo, all the oils were still included into the "extra-virgin" category, and those deriving from whole olives picked at the lowest MI showed the best sensory characteristics due to high fruity and well-balanced pungent and bitter tastes. PRACTICAL APPLICATION: This study could represent a helpful tool for oil-makers to improve the marketing of extra-virgin olive oils produced from cultivars with very high phenolic contents, such as Coratina, generally not adequately appreciated by consumers because of their excessive bitterness and pungent taste. These oils, when extracted from whole olives, are generally consumed after a certain period of time (at least 6 mo) during which a decrease in the phenolic content occurs. The results of the present work demonstrate that oils extracted from olives picked at low maturation index can be marketed immediately after production if subjected to stoning and malaxed for a short time. This procedure allows to adjust the phenolic content and to obtain a high flavor and a well-balanced taste.

Short communication: Relationships between milk quality and acidification in the production of table Mozzarella without starters.

The effect of some quality parameters of the milk (refrigeration time, pH, protein, and fat/protein ratio) on the extent of acidification in the production technology of table Mozzarella without starters was investigated. A screening phase carried out at the laboratory level demonstrated that variations of the milk characteristics require different levels of acidification to keep constant the quality of the cheese. The elaboration of the data collected throughout the successive experimentation on industrial scale allowed us to find a mathematical model to describe the relationships between the pH of the curd at stretching time and the milk characteristics, of which the protein concentration and the refrigeration time play the main roles.

Occurrence of beta-casein fragments in cold-stored and curdled river buffalo (Bubalus bubalis L.) milk.

The safeguard of river buffalo Mozzarella cheese, a Protected Designation of Origin dairy product, has prompted an analytical study to trace the milk and curd used as raw material in cheesemaking. This is to prevent the illegal use of milk or curd from different geographical areas outside of those indicated in the official production protocol. For this purpose, we studied primary proteolysis occurring in fresh and frozen milk and curd to identify a molecular marker that could indicate the raw material used. Whole casein from frozen river buffalo milk was separated using cation-exchange chromatography and sodium dodecyl sulfate-PAGE, and a protein component with an estimated molecular weight of 15.3 kDa was detected. This protein component was revealed in fresh river buffalo milk as a faint electrophoresis band, which drastically increased in intensity in refrigerated and frozen milk as well as in curd and was found to be associated with beta-CN through hydrophobic interaction. By using matrix-assisted laser desorption/ionization-time of flight peptide mass mapping, this component was identified as the C-terminal fragment f(69-209) of beta-CN (expected molecular weight of 15,748.8 Da). beta-Casein f(69-209), originating from the early hydrolysis of Lys(68)-Ser(69) by plasmin, has no counterpart in bovine milk. The increased rate of hydrolysis by plasmin toward the cleavage site Lys(68)-Ser(69) has to be ascribed to the elevated proline content of the peptide 61-73. The favored production of beta-CN f(69-209) has also drawn attention to the complementary proteose peptone beta-CN f(1-68) that is presumed to play a physiological role in inducing milk secretion similar to that of beta-CN f(1-29). The higher in vivo and in vitro production rate, compared with gamma(1)-CN formation, indicates that beta-CN f(69-209) and its complementary fragment are candidate molecular markers to evaluate milk and curd freshness. We suggested [corrected] indirect ELISA analysis based on the determination of remaining nonhydrolyzed beta-CN to perform a quantitative evaluation of proteolysis.

Molecular identification and typing of natural whey starter cultures and microbiological and compositional properties of related traditional Mozzarella cheeses.

Cell numbers of presumptive lactic acid bacteria varied markedly between 7 natural whey starter cultures (NWSC) used for producing traditional cows' milk Mozzarella cheeses in the Apulia region of Southern Italy. Taxonomic identification revealed a large diversity at species level, including mesophilic and thermophilic lactobacilli, lactococci, streptococci and enterococci. Randomly Amplified Polymorphic DNA (RAPD-PCR), analysis showed the biodiversity among the strains and, for lactobacilli, some relationships with provenience of the natural starter. Cell numbers of presumptive lactic acid bacteria in the corresponding Mozzarella cheeses were similar or higher than those found in the corresponding NWSC. RAPD-PCR analyses showed that most of the strains in cheese originated from the starter. The gross composition varied markedly between the 7 Mozzarella cheeses and ranged from 53-64% moisture, 17-23% protein, 13-20% fat and 0.50-1.61% salt. The values of pH for several samples were above 6.0. As shown by urea-PAGE of the pH 4.6-insoluble nitrogen fractions, cheese samples were characterized by differences in alpha(S1)- and beta-casein hydrolysis. Cheeses also differed with respect to secondary proteolysis as shown by Principal Component Analysis (PCA) of data from RP-HPLC of the pH 4.6-soluble, pH 4.6-70% ethanol-soluble and 70% ethanol-insoluble nitrogen fractions. These differences were attributed to the different microbial composition of the NWSC. Strain selection and optimization of a protocol for producing a natural whey starter culture to be used by dairy factories of the Apulia region appears to be a pre-requisite to standardize the major traits distinguishing this cheese variety.

[Atypical metastases from gastric carcinoma. A primary and isolated case localized in the abdominal wall]. / In tema di metastasi atipiche da carcinoma gastrico. Su un caso primitivo ed isolato localizzato alla parete addominale.

The authors report a rare case of gastric carcinoma metastasis localised in the anterior abdominal wall which was an isolated and primary sign of disease. After having underlined the rarity of the case, there is a brief discussion of aspects related to the spread of primary tumours in relation to other cases reported over the past decade; the most important reports of atypical secondary localisations are listed.

[Carcinoma of the cecum: is early diagnosis possible?]. / Il carcinoma del cieco: è possibile una diagnosi precoce?

Between 1982 and 1989, a consecutive series of 34 patients with cancer of the cecum were operated in our hospital. Overall, at 5 years the survival was 50%. We did not notice any correlation among symptom duration and survival. Patients admitted in the asymptomatic phase had the best survival rate (66.6%). Our study suggest that only neoplasm diagnosed in asymptomatic patients and treated promptly are related to a good survival rate. We, therefore, emphasize the importance and the necessity to investigate all patients with a suspected cecum cancer by means of colonoscopy.

[An unusual case of hydatid cyst with adrenal localization]. / Su un caso eccezionale di cisti idatidea a localizzazione surrenalica.

The paper reports a case of hydatid cyst with a primary adrenal location. Having underlined the rarity of this finding and reviewed the literature on the topic, the Authors discuss the epidemiological, pathogenetic and clinico-diagnostic aspects. In conclusion they underline the difficulty of arriving at a precise diagnosis prior to surgery in spite of the fact that new imaging diagnostic methods enable diagnostic hypotheses regarding this pathology to be advanced.

[Extra-ocular sebaceous carcinoma: report of a case]. / Il carcinoma sebaceo extra-oculare: a proposito di un caso.

The Authors streamed the rarity of this pathological entity for which the histologic diagnosis is even very difficult. Treatment is based on a wide surgical excision; the value of postoperative irradiation and chemotherapy is discussed. The prognosis is poor.