Inactivation of spores of Bacillus cereus in cheese by high hydrostatic pressure with the addition of nisin or lysozyme

The objective of this work was to study high hydrostatic pressure (HHP) inactivation of spores of Bacillus cereus ATCC 9139 inoculated in model cheeses made of raw milk, together with the effects of the addition of nisin or lysozyme. The concentration of spores in model cheeses was approximately 6-log10 cfu/g of cheese. Cheeses were vacuum packed and stored at 8 degrees C. All samples except controls were submitted to a germination cycle of 60 MPa at 30 degrees C for 210 min, to a vegetative cells destruction cycle of 300 or 400 MPa at 30 degrees C for 15 min, or to both treatments. Bacillus cereus counts were measured 24 h and 15 d after HHP treatment. The combination of both cycles improved the efficiency of the whole treatment. When the second pressure-cycle was of 400 MPa, the highest inactivation (2.4 +/- 0.1 log10 cfu/g) was obtained with the presence of nisin (1.56 mg/L of milk), whereas lysozyme (22.4 mg/L of milk) did not increase sensitivity of the spores to HHP. For nisin (0.05 and 1.56 mg/L of milk), no significant differences were found between counts at 24 h and 15 d after treatment. Considering that mesophilic spore counts usually range from 2.6 to 3.0 log10 cfu/ml in raw milk, HHP at mild temperatures with the addition of nisin may be useful for improving safety and preservation of soft curd cheeses made from raw milk.     

Determination of fatty acids in the subcutaneous fat of Iberian breed swine by near infrared spectroscopy (NIRS) with a fibre-optic probe

A near infrared spectrometer equipped with a standard 1210/210 bundle remote reflectance fibre-optic probe, with a 5×5 cm quartz window, was used for the determination of fatty acids in the subcutaneous fat of Iberian pigs. A comparative study was made of the determination of fatty acids (C14:0, C16:0, C18:0, C18:1, C18:2, C18:3, C20: 1, Σpolyunsaturated, Σmonounsaturated and Σsaturated) in samples of subcutaneous fat from Iberian pigs by direct application of the fibre-optic probe on samples of whole subcutaneous fat and with cam-lock cups, assessing extracts of total lipids with diethyl ether. The regression method employed was modified partial least squares (MPLS). Calibration of 157 samples, using the fibre optic probe, allowed determination of fatty acids in the following ranges: C14:0 (0.78-1.77), C16:0 (15.87-29.74), C18:0 (4.61-15.90), C18:1 (43.50-61.27), C18:2 (2.03-13.94), C18:3 (0.13-1.14), C20:1 (0.45-2.32), Σpolyunsaturated (2.31-14.82), Σmonounsaturated (47.37-65.62), Σsaturated (22.09-47.31), with corrected standard errors of prediction SEP(C) of 0.093, 0.56, 0.67, 0.94, 0.42, 0.10, 0.20, 0.46, 0.94, 0.83, respectively. The robustness of the method using the fibre-optic probe was tested in a slaughterhouse using 23 samples for external validation, giving multiple correlation coefficients (RSQ) for C14:0, C16:0, C18:0, C18:1, C18:2, C18:3 C20:1, Σpolyunsaturated, Σmonounsaturated, Σsaturated acids of 0.72, 0.94, 0.72, 0.79, 0.88, 0.55, 0.17, 0.88, 0.74, and 0.90, respectively, and a corrected standard error of prediction [SEP(C)] for these acids (%) of 0.11, 0.60, 0.84, 1.20, 0.77, 0.11, 0.30, 0.76, 1.21, and 1.18, respectively.     

Spectroscopic studies of the biosorption of gold(III) by dealginated seaweed waste

Gold biosorption by dealginated seaweed waste has been studied to elucidate the mechanisms of metal uptake from solution. Dealginated seaweed was able to retain up to 1 mmol g(-1) of Au from solution at pH 3. FT-IR showed the presence of carboxylate groups on the surface of the biosorbent; however, the changes observed for the Au-bound samples suggested very little sorption to the carboxyl moieties. Colloidal Au formed on the surface of dealginated seaweed by reduction of Au(III) to Au(0) was observed using ESEM and four different types of particles were clearly identified. The Au distribution matched closely that obtained for S atoms indicating a possible link between these elements. EXAFS measurements showed that colloidal Au is present on the surface of the biosorbent. Evidence of gold reduction from Au(III) to Au(I) and Au(0) was also confirmed by the measured bond distances characteristic of the metal. The coordination number obtained by EXAFS indicated that approximately 75% of the Au on the sample was present in the colloidal form and the remaining Au was bound to S as nearest neighbor. The proposed mechanisms for Au removal from solution are reduction of Au species by components on the surface of the biosorbent to form colloidal metal followed by retention of the ionic Au(I) species at the sulfur containing sites. The results show that dealginated seaweed can be used for the cleanup of gold-containing effluents.     

Partition of fish pheromones between water and aggregates of humic acids. Consequences for sexual signaling

High-molecular-weight suspended organic matter of soil and aquatic origins competes with water for the dissolution of relatively water insoluble organic substances. The same happens with microalgae and other organisms present in natural waters. Several pheromones, which play a specific role in the reproductive cycle of fish, are secreted to the water and are generally, if not always, molecules with hydrophobic or amphiphilic characteristics. The natural tendency of these pheromones to dissolve in suspended or deposited organic matter may cause their signaling function to be adversely affected. In this work we study the partition constants between water and organic reservoirs in suspension, Kh, of two fish pheromones, 4-pregnene-17 alpha, 20beta-diol-3-one (17,20beta-P) and prostaglandin F2alpha (PGF2alpha), and also of 4-pregnene-11beta,21-diol-3,20-dione 21-sulfate (21-P-sulfate), used as a proxy for sulfated steroid pheromones. Two types of organic reservoirs are employed: aggregates of suspended humic substances and negatively charged phospholipid vesicles. We find that the three compounds have high affinities for both types of aggregates. However, 17,20beta-P, with pKh = 4.4 +/- 0.2, is the only one for which we may predict a significant decrease in availability in consideration of the normal content of dissolved organic matter in natural waters. Also to be considered is the fact that a relatively large amount of pheromones is retained and may be released at an inopportune moment. How significant these phenomena are in nature is not as yet clear, considering the variety of habitats in which fish spawn and the little that is known about the nature and mode of action of pheromones.     

Temporal Release of Flavor Compounds from Low-fat and High-fat Ice Cream During Eating

ABSTRACT: Fat controls the intensity and the release rate of volatile compounds because of its solvent property. Our objective was to understand the effect of fat on the flavor release behavior exhibited while ice cream is eaten. The temporal flavor profiles of ice creams, which contained various fat levels and were spiked with known amounts of selected volatiles, were determined by a temporal modified purge and trap and time-intensity analyses. Stale and cherry flavors were released faster and at higher intensities as fat content decreased. Fat had the opposite effect on vanilla flavor release. Temporal modified purge and trap analysis was an effective method to observe the release of some flavor compounds.     

Antimicrobial activity of bovine β-lactoglobulin against mastitis-causing bacteria

Bovine mastitis is one of the most economically deleterious diseases affecting dairy herds and results from an infection of the udder by pathogenic microorganisms such as Staphylococcus aureus, Streptococcus uberis, and Escherichia coli. The mammary gland is capable of preventing and combating bacterial infection by means of a complex network of innate and adaptive immune mechanisms. Lactoferrin is an 86-kDa protein with antibacterial activity that plays a role in the mammary gland's defense against infection. β-Lactoglobulin (β-LG) is an 18-kDa protein that is present in most mammals but is notably absent in humans, rodents, and lagomorphs. Different genetic variants of this protein exist, with β-LG A and β-LG B being the most common. In spite of being well studied, the biological function of β-LG is not thoroughly understood, and most noticeably, no reports exist on the effects of the native protein on bacterial growth. Hence, the objective of this study was to assess the potential antibacterial activity of β-LG against mastitis agents. To do this, we purified β-LG from normal bovine milk using a mild, nondenaturing method and performed in vitro growth inhibition assays with Staph. aureus, E. coli, and Strep. uberis. β-Lactoglobulin inhibited the growth of Staph. aureus and Strep. uberis but had no effect on E. coli. The antimicrobial activity against Staph. aureus and Strep. uberis was concentration dependent and was elicited by the intact protein because Tricine-sodium dodecyl sulfate-PAGE and analytical gel filtration chromatography did not reveal the presence of short degradation peptides. Analysis of the genetic variants of β-LG showed that β-LG A has higher inhibitory activity against Staph. aureus and Strep. uberis than β-LG B. Coincubation of β-LG and lactoferrin resulted in an augmented antibacterial activity against Staph. aureus, suggesting an additive effect of the proteins. This result, along with the proteins’ complementary spectrum of action, suggests that β-LG and lactoferrin may complement each other in the mammary gland's defenses against bacterial infection.     

Short communication: Effect of subclinical mastitis on proteolysis in ovine milk

The aim of this work was to evaluate the effect of intramammary infection (IMI) on the endogenous proteolysis of milk. Four control checks were carried out in the half-udder milk of 10 ewes that acquired unilateral subclinical mastitis. Two of these checks were conducted before the infection was established and 2 after. Ten healthy ewes were tested as a control group. The presence of a subclinical IMI involved an increase of the products of casein hydrolysis, the proteose-peptone (p-p) fraction and minor (m) caseins, and a decrease of β-casein. As a result, a significant increase in the proteolysis index (PI), calculated as the ratio of m-casein to the sum of caseins (α + β + κ), took place. α-Casein and κ-casein were not significantly affected by IMI. Correlations confirmed the scenario: log₁₀ of somatic cell count (SCC) was positively correlated with p-p content and negatively with β-casein, whereas log₁₀ SCC was not correlated with α-casein or κ-casein. On the other hand, p-p content was positively correlated with m-casein and PI and negatively with β-casein, but no correlation was detected between p-p content and α- or κ-casein. Furthermore, between casein fractions, m-casein was only significantly correlated with β-casein. These results suggest that use of indices of proteolysis of caseins such as p-p, m-casein, and PI, could be applied together with SCC to evaluate the cheese-making quality of milk.     

Antioxidant activity of several marine skin gelatins

Gelatins obtained from the skins of tuna and halibut, and from the tunics of jumbo flying squid were hydrolysed by Alcalase to produce antioxidant peptides. Hydrolysis yielded an increase in the antioxidant capacity of gelatins of around two-fold when measured by the Fe reducing capacity (FRAP) method and even more when measured by the ABTS radical scavenging method. When both squid and tuna gelatins were hydrolysed with different enzymes (collagenase, trypsin, pepsin), Alcalase and pepsin gave the hydrolysates with the highest and lowest ABTS radical scavenging ability, respectively. FRAP assay showed that the squid hydrolysates prepared using Alcalase were the most effective in reducing ferric ions, whereas trypsin gave rise to the tuna hydrolysates with the highest iron reducing ability. When the amino acid composition of the gelatins was related to the antioxidant properties, ABTS radical scavenging was observed to be negatively correlated to the total content of hydrophobic amino acids and imino acids in all the samples, while Fe reducing power (FRAP) was strongly correlated with Hyl and degree of hydroxylation.     

Chemical composition, mineral content and antioxidant activity of Verbena officinalis L.

Aqueous and hydroalcoholic extracts from Verbena officinalis L. were obtained and characterised. The analysis by HPLC-DAD and LC-MS allowed the detection and identification of three iridoids, fifteen flavonoids and four phenolic acid derivatives. Four flavonoids, scutellarein 7-diglucuronide (9), scutellarein 7-glucuronide (13), pedalitin 6-galactoside (15) and scutellarein 7-glucoside (19) are reported for the first time from this plant. In addition, three new flavonoids have been isolated: scutellarein 7-O-(2-O-feruloyl)-diglucuronide (5), pedalitin 6-O-diglucuronide (6) and pedalitin 6-O-(2-O-feruloyl)-diglucuronide (13). To our knowledge, these flavonoids have not been reported as natural products. Both extracts showed significant antioxidant activity using three in vitro model systems and the results have been correlated with total phenolic and total flavonoid contents. The results have allowed establishing an important relation structure-activity and significant correlations have also been found between the mineral content and the flavonoids present in both extracts.     

Extracellular polysaccharides production by Arthrobacter viscosus

In this study the production of extracellular polysaccharides by the non-pathogenic soil bacteria Arthrobacter viscosus has been investigated. Different variables affecting extracellular polysaccharide production such as the carbon source (glucose or xylose), the agitation speed and the pH have been analysed.

In a first stage, experiments in shaken conical flasks (250 ml), containing 50 ml of culture medium, were carried out. Using xylose (25 g/l) as the carbon source at an initial pH 8 improved the extracellular polysaccharides levels obtained.

In a second stage, the experiments were scaling in bioreactors. Cultivation was carried out in discontinuous mode and with/without pH control. Polysaccharide production reached a maximum of 10 g of crude product per litre of growth medium after 14 days and the relationship between product formation and cell growth of A. viscosus is 2.7 g polysaccharide per gram biomass. This production was obtained at the optimal conditions determined with pH control at pH 7, xylose as carbon source (25 g/l) and an agitation rate of 800 rpm.