Formation of the aroma of a raw goat milk cheese during maturation analysed by SPME–GC–MS

The volatile profile of the Spanish goat raw milk cheese of the protected designation of origin (PDO) “Queso Ibores” was studied at four stages of maturation (day 1, 30, 60, and 90) by the method of solid-phase micro-extraction–gas chromatography–mass spectrometry (SPME–GC–MS) to determinate the characteristic volatile compounds of this cheese and to know the changes in the volatile profile of this cheese during maturation. According to the PDO, Ibores cheese aroma varies between sweet and mild and it has a strong taste, slightly tart. A total of 64 compounds were detected: 14 acids, 18 alcohols, 13 esters, 6 ketones and 13 compounds which could not be classified in these groups. Carboxylic acids were the most abundant volatile compounds in the headspace of Ibores cheese. Content of volatile compounds was significantly modified (P < 0.05) during ripening. The relative total amounts of acids, esters and ketones increased during the first 60 days of maturation. The most characteristic compounds of Ibores cheese aroma were butanoic, hexanoic and octanoic acids, some alcohols (2-butanol and 2-heptanol), ethyl esters of hexanoic and butanoic acids, some methyl ketones (2-butanone, 2-pentanone and 2-heptanone) and δ-decalactone.     

High occurrence of Helicobacter pylori in raw goat, sheep and cow milk inferred by glmM gene: a risk of food-borne infection?

Helicobacter pylori is an organism widespread in humans and sometimes responsible for serious illnesses, such as gastric and duodenal ulcers, MALToma and even gastric cancer. It has been hypothesized that the infection route by H. pylori involves multiple pathways including food-borne transmission, as the microorganism has been detected from foods such as sheep and cow milk. This work reports the results of a survey conducted in order to investigate the presence of H. pylori in raw goat, sheep and cow milk produced in Southern Italy, employing a Nested Polymerase Chain Reaction (Nested-PCR) assay for the detection of the phosphoglucosamine mutase gene (glmM), as screening method followed by conventional bacteriological isolation. Out of the 400 raw milk samples examined, 139 (34.7%) resulted positive for the presence of glmM gene, but no strains were isolated. In this work H. pylori DNA has been firstly detected from 41 (25.6%) raw goat milk samples. The results deserve further investigations on the contamination source/s of the milk samples and on the major impact that it may have on consumers.     

Monitoring and identification of bacteria associated with safety concerns in the manufacture of São Jorge, a Portuguese traditional cheese from raw cow's milk

The aim of this study was to assess the hygienic quality of raw milk used in the manufacture of S?o Jorge, a Protected Denomination of Origin Portuguese semihard cheese, as well as to ascertain the sanitary conditions prevailing during its processing. Viable counts of Enterobacteriaceae and Micrococcaceae were accordingly obtained, pertaining to 21 independent batches (including samples of raw milk, curd, and cheeses after 1, 3, and 4 months of ripening), from 7 dairy farms. Standard plate counts (log CFU per milliliter or per gram) ranged from 6.1 to 8.6 in raw milk, whereas they ranged from 7.0 to 8.0 in 4-month-old cheeses. Viable counts of Enterobacteriaceae ranged between 5.9 and 7.0 in raw milk and between 0.0 and 1.3 in 4-month-old cheeses. Species identified within this family encompassed Klebsiella oxytoca, Klebsiella cloacae, Klebsiella pneumoniae, Enterobacter sakazakii, and Escherichia coli; Klebsiella ornithinolytica, Klebsiella terrigena, and Serratia odorifera were detected only in raw milk. No Salmonella whatsoever could be detected in any of the samples. Viable counts of Micrococcaceae ranged between 4.7 and 5.9 and between 1.3 and 3.3 in raw milk and 4-month-old cheeses, respectively. Species identified within this family encompassed Staphylococcus sciuri, Staphylococcus epidermidis, Staphylococcus saprophyticus (which was found mainly in ripened cheeses), and Staphylococcus aureus (which was not detected in 4-month-old cheeses). Accompanying physicochemical analyses included determination of moisture, salt, and pH. Statistical analyses revealed a negative correlation between salt content and viable numbers of Enterobacteriaceae in cheese, whereas in the case of Micrococcaceae, a more negative correlation was found between viable numbers and moisture content than between viable numbers and pH. The results of our study indicate, in general, poor milk handling conditions in all farms, given that the indicators total mesophile and Enterobacteriaceae counts were high, between 100- and 1,000-fold those enforced by international standards pertaining to the matrices in question. However, by the time of regular consumption (i.e., after 4 months of ripening), S?o Jorge cheeses exhibit low levels of contamination by Enterobacteriaceae and S. aureus, as well as absence of Salmonella.     

The tentative recognition of psychrotrophic Gram-negative bacteria in 48 h by their surface growth at 10°C

Employing 153 species and strains of Gram-negative bacteria representing 20 genera, 79 produced visible growth on brain heart infusion agar (BHIA) at 7°C in 10 days and thus conformed to the definition of psychrotrophs. All but 5 of the 79 (93.7%) grew on BHIA plates at 10°C in 2 days. At 12°C, 13 psychrotrophs did not grow in 24 h while in 2 days 28 nonpsychrotrophs grew. Surface plating for colony enumeration generally required 3 days, and pour plating 4 days for countable colonies. It is suggested that those bacteria that grow rapidly at 7°C but not at 40°C be designated stenopsychrotrophs while those that can grow at 40°C or above and generally more slowly at 7°C be designated eurypsychrotrophs. The use of 10°C for 2 days may be used to make a rapid and tentative designation of Gram-negative psychrotrophs when turbid broth cultures are streaked on BHIA plates.     

Determination of sulbactam in raw bovine milk by isotope dilution–ultra-high-performance liquid chromatography–tandem mass spectrometry

We developed a sensitive and selective isotope dilution ultra-high-performance liquid chromatography–tandem mass spectrometry (UPLC-MS/MS) method for the determination of sulbactam residue in raw bovine milk. Sulbactam and internal standard, sulbactam-d₅, were extracted from raw bovine milk via liquid-liquid extraction and enriched with strong anion exchange solid-phase extraction cartridges and finally analyzed by using UPLC-MS/MS with multiple reaction monitoring mode. The method was validated according to European regulations. The calibration curve showed good linearity, with a correlation coefficient of 0.9998. Decision limit and detection capability of sulbactam were determined by matrix calibration curve and were 0.0445 and 0.0517 μg/L, respectively. The recoveries of sulbactam in fortified raw bovine milk ranged from 72.1 to 91.5%, with the intra- and interday relative standard deviations ranging from 3.0 to 18.9%. Furthermore, the developed method was applied to analyzing real raw bovine milk samples collected from dairy farms in Beijing, China. Sulbactam was not determined in all samples. The proposed method could ultimately serve as a methodological foundation for the determination of sulbactam in different types of raw milk and dairy products.     

Simple membrane filtration method for estimating numbers of Paenibacillus spp. spores in raw milk,using β-galactosidase activity as a selection criterion

Paenibacillus spp. are spore-forming bacteria that adversely affect the quality of dairy products. There is currently no appropriate method for enumerating Paenibacillus spp. spores. We developed a simple membrane filtration method to enumerate Paenibacillus spp. spores in raw milk, using β-galactosidase activity as a selection criterion. Although Paenibacillus spp. spores are relatively small, use of a membrane filter with 0.65-μm pore size allowed us to easily filter raw milk with sufficient recovery. The membrane was put on plates containing X-gal, and detection of β-galactosidase-positive colonies enabled selective enumeration of Paenibacillus spp. spores. We investigated Paenibacillus spp. spore levels in raw milk from six different areas in the Tokachi region, Hokkaido, Japan over 1 year. There were ≤10 spores 100 mL⁻¹ throughout the year, with no significant differences between areas or seasons. Paenibacillus amylolyticus and Paenibacillus odorifer were the predominant species, accounting for 50.6% and 27.4% of the total spores, respectively.     

Goat milk allergenicity as a function of αs₁-casein genetic polymorphism

Cow milk allergy is the most frequent allergy in the first years of life. Milk from other mammalian species has been suggested as a possible nutritional alternative to cow milk, but in several cases, the clinical studies showed a high risk of cross-reactivity with cow milk. In the goat species, α_S1-casein (α_S1-CN), coded by the CSN1S1 gene, is characterized by extensive qualitative and quantitative polymorphisms. Some alleles are associated with null (i.e., CSN1S1*0₁) or reduced (i.e., CSN1S1*F) expression of the specific protein. The aim of this work was to obtain new information on goat milk and to evaluate its suitability for allergic subjects, depending on the genetic variation at α_s1-CN. Individual milk samples from 25 goats with different CSN1S1 genotypes were analyzed by sodium dodecyl sulfate PAGE and immunoblotting, using monoclonal antibodies specific for bovine α-CN and sera from children allergic to cow milk. A lower reaction was observed to 2 goat milk samples characterized by the CSN1S1* 0₁0₁ and 0₁F genotypes. Moreover, a fresh food skin prick test, carried out on 6 allergic children, showed the lack of positive reaction to the 0₁0₁ milk sample and only one weak reactivity to the 0₁F sample. The risk of cross-reactivity between cow and goat milk proteins suggests the need for caution before using goat milk for infant formulas. However, we hypothesize that it can be used successfully in the preparation of modified formulas for selected groups of allergic patients. The importance of taking the individual goat CN genetic variation into account in further experimental studies is evident from the results of the present work.     

Could modifications of processing parameters enhance the growth and selection of lactic acid bacteria in cold-smoked salmon to improve preservation by natural means?

Several smoking conditions were examined with the objective of enhancing the numbers of lactic acid bacteria (LAB) by natural means in vacuum-packaged cold-smoked salmon during 21 days of storage at 5 degrees C. Three combinations of salting, drying, and smoking were used: (i) dry salting x time of salting (2 or 6 h); (ii) wet salting (6 h) x dry salting (6 h) x with or without sugar; and (iii) wet salting (6 h) x dry salting (6 h) x different times of smoking (2 or 6 h of drying and 2 or 6 h of smoking). Two batches were processed for each set of conditions. Determinations of pH and salt content in the water phase were carried out for products in each treatment. Microbiological analyses (total viable count, total LAB, Lactobacillus spp., and Enterobacteriaceae) also were conducted at the beginning of storage (t0) and after 21 days of refrigerated storage (tl). There were differential increases in total LAB and lactobacilli during the storage period according to the treatment performed. The most effective treatment to enhance LAB growth was 6 h of dry salting with sugar, 6 h of drying, and 2 h of smoking. These salting-drying-smoking conditions also selected the LAB as the dominant flora at the end of the storage period. The LAB promoted by these processing parameters seem to be potentially useful protective cultures because of their anti-Listeria activity. From the results of this research, we conclude that it is possible to enhance the growth of LAB in general and that of inhibitory strains in particular by suitable choices of processing parameters.     

Visible and near-infrared spectroscopic analysis of raw milk for cow health monitoring: reflectance or transmittance?

The composition of produced milk has great value for the dairy farmer. It determines the economic value of the milk and provides valuable information about the metabolism of the corresponding cow. Therefore, online measurement of milk components during milking 2 or more times per day would provide knowledge about the current health and nutritional status of each cow individually. This information provides a solid basis for optimizing cow management. The potential of visible and near-infrared (Vis/NIR) spectroscopy for predicting the fat, crude protein, lactose, and urea content of raw milk online during milking was, therefore, investigated in this study. Two measurement modes (reflectance and transmittance) and different wavelength ranges for Vis/NIR spectroscopy were evaluated and their ability to measure the milk composition online was compared. The Vis/NIR reflectance measurements allowed for very accurate monitoring of the fat and crude protein content in raw milk (R² > 0.95), but resulted in poor lactose predictions (R² < 0.75). In contrast, Vis/NIR transmittance spectra of the milk samples gave accurate fat and crude protein predictions (R² > 0.90) and useful lactose predictions (R² = 0.88). Neither Vis/NIR reflectance nor transmittance spectroscopy lead to an acceptable prediction of the milk urea content. Transmittance spectroscopy can thus be used to predict the 3 major milk components, but with lower accuracy for fat and crude protein than the reflectance mode. Moreover, the small sample thickness (1 mm) required for NIR transmittance measurement considerably complicates its online use.     

Could predicting fatty acid profile by mid-infrared reflectance spectroscopy be used as a method to increase the value added by milk production chains?

The aims of this work were (1) to develop prediction equations from mid-infrared spectroscopy (MIRS) to establish a detailed fatty acid (FA) composition of milk; (2) to propose a milk FA index, utilizing MIRS-developed equations, in which the precision of the FA-prediction equations is taken into account to increase the value of milk; and (3) to show application examples. A total of 651 bulk cow milk samples were collected from 245 commercial farms in northwest Italy. The results of the 651 gas chromatography analyses were used to establish (421 samples) and to validate (230 samples) the outcomes of the FA composition prediction that had been obtained by MIRS. A class-based approach, in which the obtained MIRS equations were used, was proposed to define a milk classification. The method provides a numerical index [milk FA index (MFAI)] that allows a premium price to be quantified to increase the value of a favorable FA profile of milk. Ten FA were selected to calculate MFAI, according to their relevance for human health and potential cheese sensory properties, and animal welfare and environmental sustainability were also considered. These factors were selected as dimensions of MFAI. A statistical analysis and expert judgment aggregation were performed on the selected FA by weighting the FA and normalizing the dimensions to reduce redundancy. A class approach was applied, using the precision of the MIRS equations to establish the classes. The median FA concentration of the data set was set as a reference value of class 0. The width, number, and limits of classes above and below the median were calculated using the 95% confidence level of the standard error of prediction, corrected with the bias of each FA. A progressive number and a positive or negative sign were assigned to each FA class above or below the median according to their role in the above mentioned dimensions. The sum of the numbers of each class, associated with its sign for each FA, was used to generate MFAI. The MFAI was applied to dairy farms characterized by different feeding strategies, all of which deliver milk to a commercial dairy plant. The MFAI values ranged from 0.7 to 4.2, and large variations, which depended on the cows' diet and forage quality, were observed for each feeding system. The proposed method has been found to be flexible and adaptable to several contexts on both intensive and extensive dairy farms.     

Technological characterization of lactic acid bacteria isolated from Armada cheese (a Spanish goats’ milk cheese)

Thirty-one strains of lactic acid bacteria (LAB) isolated from Armada cheese, Sobado variety, (eight strains of Lactococcus lactis subsp. lactis, four strains of Lactococcus lactis subsp. cremoris, two strains of L. lactis subsp. lactis biovar. diacetylactis, two strains of Leuconostoc mesenteroides subsp. mesenteroides, two strains of Leuconostoc mesenteroides subsp. dextranicum, five strains of Lactobacillus plantarum, six strains of Lactobacillus casei subsp. casei and two strains of Lactobacillus brevis) were screened for their acidifying capacity and enzymatic activity, that included the rapid API-ZYM system, the proteolytic activity, the amino-, di-, and carboxypeptidase activity and the caseinolytic activity. The strains of L. lactis subsp. lactis exhibited the highest acidifying and proteolytic activity. Lipase and esterase activity was practically non-existent for lactococci and lactobacilli; a certain esterase activity was observed among leuconostoc. The highest aminopeptidase activity was demonstrated by the cell-free extract (CFE) of some strains of L. plantarum, L. casei subsp. casei and L. mesenteroides subsp. dextranicum. The CFEs of L. lactis subsp. cremoris and L. lactis subsp. lactis possessed carboxypeptidase and dipeptidase activities, at levels depending on the strain. Appreciable caseinolytic activity was detected for the CFE of L. plantarum and those some lactococci.     

Utilization of sprout‐damaged wheat as raw material for sour dough pre‐ferments with mixed cultures of lactic and propionic acid bacteria

The utilization of sprout‐damaged wheat flour in pre‐fermentation for sour dough bread‐baking increased lactic acid formation and titratable acidity several fold in comparison to the use of baker's wheat flour, and also resulted in a lactic acid to acetic acid molar ratio acceptable for sour dough bread. Further, enzymic treatment of the sprout‐damaged wheat flour with malted grain flour appared to be necessary for a properly balanced acid formation although the α‐amylase activity of sprout‐damaged wheat flour was quite high. The accumulation of lactic acid in the pre‐ferment decreased in the presence of yeast, and thus the molar ratio of lactic to acetic acid could be adjusted suitable for sour bread‐baking even with homofermentative lactic acid bacteria. Finally, when using mixed culture pre‐ferments of Lactobacillus brevis and Propionibacterium jensenii, at least 0.1% of propionic acid (based on flour weight), sufficient to prevent mold growth in the final bread, was obtained.     

Determination of Nϵ-carboxymethyllysine in milk products by a modified reversed-phase HPLC method

A modified reversed-phase-HPLC method with o-phthalaldehyde pre-column-derivatisation for determination of N^ϵ-carboxymethyllysine in food samples is presented. It is shown, that the method has to be modified if applied to milk products, including specific modifications in sample preparation and chromatographic separation conditions. The increased selectivity of a double endcapped RP 18 phase is necessary for reliable separation of N^ϵ-carboxymethyllysine in hydrolysates of complex products like cheese. With a detection limit of 0.5 pMol the method shows high sensitivity and a very good reproducibility (s=2.81%). In total, several different milk products (n=50) as well as fresh, processed and ripened cheese samples (n=50) were analysed. The highest amounts of N^ϵ-carboxymethyllysine were found in a whey cheese (1016 mg/kg protein), evaporated milk (1691 mg CML/kg protein), coffee cream (613 mg CML/kg protein) and cocoa milk (413 mg CML/kg protein). N^ϵ-carboxymethyllysine could not be detected in UHT milk, fresh, processed and ripened cheese. The results show that N^ϵ-carboxymethyllysine can give valuable information on lysine damage in severely heat-treated milk products and in products, with added sugar, pre-damaged constituents or stabilising agents. ©     

Determination of the bond strength of treated wood strands embedded in a cement matrix by means of a pull-out test

Effects of chemical treatment of wood on the bonding strength of Norway spruce strands (Picea abies Karst.) embedded in a??cement matrix were investigated by means of a??pull-out test. Strands of varying thickness were used whereas a??strong influence of thickness on bonding strength could be observed. Using thick strands (800?μm) showed negative effect on bonding strength which is due to the swelling effect. The treatment of thin strands (400?μm) showed that using wet strands (～?90% moisture content) leads to the best results compared to untreated dry (12%) strands. By addition of setting accelerators (ammonium chloride) bonding strength of wet strands was even improved. Furthermore, effects of hot water and sodium hydroxide extraction as well as sodium silicate treatment were examined.     

Agar-immobilized basil–lactic acid bacteria bioproducts as goat milk taste-masking agents and natural preservatives for the production of unripened goat cheese

Goat milk cheeses have become popular recently; however, many consumers do not choose these products because they have specific sensory properties that are not acceptable to all consumers and the shelf life of the cheese is short. The concept of this work was to increase overall acceptability and shelf life of unripened goat milk cheese by using Ocimum basilicum and lactic acid bacteria (Lactobacillus plantarum LUHS135, Lactobacillus paracasei LUHS244, Pediococcus pentosaceus LUHS100, Pediococcus acidilactici LUHS29, and Lactobacillus brevis LUHS140) bioproducts (basil-LAB) immobilized in agar. A basil-LAB bioproduct could be a promising multifunctional ingredient for cheese manufacturing because it has a low pH, high LAB count, and high total phenolic compound content (after fermentation pH decreased by 25.4%, LAB count averaged 7.2 log₁₀ cfu/g, and total phenolic compound content increased by 30.9%). Use of different LAB in the preparation of basil-LAB bioproducts had a significant influence on cheese pH and hardness, and compared with cheese samples prepared with nonfermented basil, cheese samples prepared with basil-LAB bioproducts had, on average, higher pH (by 2.6%) and lower hardness (by 36.0%), similar to the control cheese (without basil). Overall acceptability of cheese was significantly influenced by the basil-LAB bioproduct immobilization process; in all cases, cheese samples prepared with fermented and immobilized basil-LAB bioproduct had better acceptability (5 points). After 120 h of storage, cheese samples prepared with basil-LAB bioproducts fermented with LUHS135, LUHS244 and LUHS140, no enterobacteria were found, and we detected strong negative and moderate negative correlations, respectively, of LAB count with enterobacteria count and yeast/mold count (r = ?0.7939 and r = ?0.4495, respectively). Finally, immobilization increased LAB viability in fresh goat milk cheese, which led to a reduction in enterobacteria and mold/yeast contamination during storage and an increase in overall acceptability compared with nonimmobilized basil-LAB. Therefore, basil-LAB bioproducts fermented with LUHS135, LUHS244, and LUHS140 strains can be recommended for preparing fresh goat milk cheese with extended shelf life and high acceptability.     

Isolation of β-lactoglobulin from buffalo milk and characterization of its structure changes as a function of pH and ionic strength

β-lactoglobulin (β-lg) is one of the most widely used proteins in food industry. Purification of β-lg from buffalo milk while maintaining its native structure is of practical influence to the industry. This research demonstrated that β-lg can be separated efficiently from whey using 70 g kg^??1 NaCl at pH 2.0, which showed higher yield and activity of β-lg. It was found that the presence of NaCl or the changing of pH had no influence on the secondary structure of β-lg while they significant influenced the tertiary structure of β-lg. More hydrophobic groups were exposed with increasing NaCl concentration. Presence of low concentration of NaCl inhibited the exposure of Trp while the exposure of Trp was increased in the presence of high concentration of NaCl. In contrast, pH changes did not influence the exposure of Trp while the decrease of pH in the range of 7.5–5.5 decreased the exposure of hydrophobic groups to the hydrophilic environment.     

Equivalent processing for pasteurization of a pineapple juice–coconut milk blend by selected nonthermal technologies

Identifying equivalent processing conditions is critical for the relevant comparison of food quality attributes. This study investigates equivalent processes for at least 5-log reduction of Escherichia coli and Listeria innocua in pineapple juice–coconut milk (PC) blends by high-pressure processing (HPP), pulsed electric fields (PEF), and ultrasound (US) either alone or combined with other preservation factors (pH, nisin, and/or heat). The two blends (pH 4 and 5) and coconut milk (pH 7) as a reference were subjected to HPP at 300–600 MPa, 20°C for 0.5–30 min; PEF at an electric field strength of 10–21 kV/cm, 40°C for 24 µs; and US at 120 µm amplitude, 25 or 45°C for 6 or 10 min. At least a 5-log reduction of E. coli was achieved at pH 4 by HPP at 400 MPa, 20°C for 1 min; PEF at 21 kV/cm, 235 Hz, 40°C for 24 µs; and US at 120 µm, 45°C for 6 min. As L. innocua showed greater resistance, a synergistic lethal effect was provided at pH 4 by HPP with 75 ppm nisin at 600 MPa, 20°C for 5 min; PEF with 50 ppm nisin at 18 kV/cm, 588 Hz, 40°C for 24 µs; and US at 45°C, 120 µm for 10 min. The total soluble solids (11.2–12.4°Bx), acidity (0.47%–0.51% citric acid), pH (3.91–4.16), and viscosity (3.55 × 10⁻³–4.0 × 10⁻³ Pa s) were not significantly affected under the identified equivalent conditions. HPP was superior to PEF and US, achieving higher ascorbic acid retention and lower color difference in PC blend compared to the untreated sample.     

Heat stability of concentrated skim milk as a function of heating time and temperature on a laboratory scale – Improved methodology and kinetic relationship

The destabilising effect of heat treatment on skim milk concentrated by reverse osmosis was shown to be strongly dependent on heating time and temperature. Heat-induced coagulation could be described as a function of heating time and temperature over a broad range of non-fat total solids concentration using the described test method. This improved test method in terms of a more instant and better visibility of coagulation effects can be used for a wide range of total solids. Heat stability strongly decreased as total solids content increased as a function of heating time and temperature. A clear correlation between heat stability of unconcentrated skim milk and concentrates thereof could be shown. In this case, the heat stability of unconcentrated skim milk can be used to estimate heat stability of concentrated skim milk as variation in milk composition of pooled bulk milk from a local dairy plant was seen to be low.