Examination of nanoparticle inactivation of Campylobacter jejuni biofilms using infrared and Raman spectroscopies.

AIMS: To investigate inactivation effect and mechanism of zinc oxide nanoparticles (ZnO NPs) activity against Campylobacter jejuni biofilms. METHODS AND RESULTS: ZnO NPs with concentrations of 0, 0·6, 1·2 and 6 mmol l(-1) were employed in antimicrobial tests against Camp. jejuni planktonic cells and biofilms. Campylobacter jejuni sessile cells in biofilms were more resistant to a low concentration of ZnO NPs when compared to planktonic cells. The ZnO NPs penetrated the extracellular polymeric substance (EPS) without damage to the EPS and directly interacted with the sessile bacterial cells, as determined using infrared spectroscopy and scanning electron microscopy. Raman spectroscopy shows alterations in quinone structures and damage to nucleic acids following Camp. jejuni treatment with ZnO NPs. The mechanism of DNA damage is most likely due to the generation of reactive oxygen species (ROS). Spectroscopic-based partial least squares regression (PLSR) models could predict the number of surviving sessile cell numbers within a bacterial biofilm (≥log 4 CFU, root mean square error of estimation <0·36) from Fourier transform infrared (FT-IR) spectral measurements. CONCLUSIONS: ZnO NPs were found to have antimicrobial activity against Camp. jejuni biofilms. ZnO NPs penetrated the biofilm EPS within 1 h without damaging it and interacted directly with sessile cells in biofilms. Alterations in the DNA/RNA bases, which are owing to the generation of ROS, appear to result in Camp. jejuni cell death. SIGNIFICANCE AND IMPACT OF THE STUDY: ZnO NPs may offer a realistic strategy to eliminate Camp. jejuni biofilms in the environment.

Evaluation of a new enrichment broth for detection of Cronobacter spp. in powdered infant formula.

The aim of this study was to investigate the potential of using Al-Holy-Rasco (AR) medium, a novel broth for detection and isolation of Cronobacter spp. in infant formula milk (IFM). The new medium's composition is generic brain heart infusion broth with the addition of 1% NaCl, 15% sucrose, and 0.80 g/liter sodium deoxycholate as selective ingredients. AR broth outperformed Enterobacteriaceae enrichment broth (EE), Enterobacter sakazakii enrichment broth (ESE), modified lauryl sulfate broth, and milk as enrichment media to stimulate the growth of a cocktail of 10 strains of Cronobacter. Additionally, AR broth significantly suppressed the growth of competing non-Cronobacter Enterobacteriaceae as compared with EE, ESE, modified lauryl sulfate broth, and milk. The recovery of desiccated Cronobacter (1 to 5,000 CFU/100 g) from powdered IFM in the presence of competing non-Cronobacter Enterobacteriaceae was determined by EE, ESE, and AR broth with 10 and 15% sucrose. AR broth with 15% sucrose outperformed all other examined broths and recovered Cronobacter from all samples tested at all Cronobacter concentrations. AR broth must be validated before it can be used for rapid detection and isolation of Cronobacter from powdered IFM.

Detection of sublethal thermal injury in Salmonella enterica serotype typhimurium and Listeria monocytogenes using Fourier transform infrared (FT-IR) spectroscopy (4000 to 600 cm(-1)).

Fourier transform infrared (FT-IR) spectroscopy (4000 to 600 cm(-1)) was utilized to detect sublethally heat-injured microorganisms: Salmonella enterica serotype Typhimurium ATCC 14028, a Gram-negative bacterium, and Listeria monocytogenes ATCC 19113, a Gram-positive bacterium. A range of heat treatments (N= 2) at 60 degrees C were evaluated: 0D (control), 2D, 4D, 6D, and 8D using a D(60 degrees C) (S. enterica serotype Typhimurium ATCC 14028 = 0.30 min, L. monocytogenes ATCC 19113 = 0.43 min). The mechanism of cell injury appeared to be different for Gram-negative and Gram-positive microbes as observed from differences in the 2nd derivative transformations and loadings plot of bacterial spectra following heat treatment. The loadings for PC1 and PC2 confirmed that the amide I and amide II bands were the major contribution to spectral variation, with relatively small contributions from C-H deformations, the antisymmetric P==O stretching modes of the phosphodiester nucleic acid backbone, and the C-O-C stretching modes of polysaccharides. Using soft independent modeling of class analogy (SIMCA), the extent of injury could be predicted correctly at least 83% of the time. Partial least squares (PLS) calibration analysis was constructed using 5 latent variables for predicting the bacterial counts for survivors of the different heat treatments and yielded a high correlation coefficient (R= 0.97 [S. enterica serotype Typhimurium] and 0.98 [L. monocytogenes]) and a standard error of prediction (SEP= 0.51 [S. enterica serotype Typhimurium] and 0.39 log(10) CFU/mL [L. monocytogenes]), indicating that the degree of heat injury could be predicted.

Monitoring quality loss of pasteurized skim milk using visible and short wavelength near-infrared spectroscopy and multivariate analysis.

Visible and short wavelength near-infrared diffuse reflectance spectroscopy (600 to 1,100 nm) was evaluated as a technique for detecting and monitoring spoilage of pasteurized skim milk at 3 storage temperatures (6, 21, and 37 degrees C) over 3 to 30 h (control, t = 0 h; n = 3). Spectra, total aerobic plate count, and pH were obtained, with a total of 60 spectra acquired per sample. Multivariate statistical procedures, including principal component analysis, soft independent modeling of class analogy, and partial least squares calibration models were developed for predicting the degree of milk spoilage. Principal component analysis showed apparent clustering and segregation of milk samples that were stored at different time intervals. Milk samples that were stored for 30 h or less at different temperatures were noticeably separated from control and distinctly clustered. Soft independent modeling of class analogy analysis could correctly classify 88 to 93% of spectra of incubated samples from control at 30 h. A partial least squares model with 5 latent variables correlating spectral features with bacterial counts and pH yielded a correlation coefficient (R = 0.99 and 0.99) and a standard error of prediction (0.34 log(10) cfu/mL and 0.031 pH unit), respectively. It may be feasible to use short wavelength near-infrared spectroscopy to detect and monitor milk spoilage rapidly and noninvasively by correlating changes in spectral features with the level of bacterial proliferation and milk spoilage.

Effect of temperature (-5 to 130 degrees C) and fiber direction on the dielectric properties of beef Semitendinosus at radio frequency and microwave frequencies.

The dielectric properties must be defined to design efficient radio frequency (RF) and microwave (MW) processes by the food manufacturers. The objective of this study was to understand how frequency, temperature, and muscle fiber orientation influence the dielectric properties. The eye of round (Semitendinosus) muscle was selected because it contains large, relatively uniform muscle cells with similar muscle fiber orientation and relatively uniform chemical composition throughout the tissue. Dielectric properties were measured using an open-ended coaxial probe technique at 27, 915, and 1800 MHz and temperatures between -5 and 130 degrees C. Power penetration depth was calculated. Since many commercially prepared, thermally processed, ready-to-eat entrees are made with frozen meat, dielectric property measurements were started from -5 degrees C. The dielectric constant and dielectric loss factors were often higher for muscle with the muscle fiber measured in a parallel orientation to the probe compared to samples of the same treatment (for example, fresh or frozen) in a perpendicular tissue orientation at the same frequency and temperature. Dielectric constant and loss values for frozen beef tended to be higher than fresh beef at the same temperature and frequency. Tissue orientation appeared to have a greater effect on dielectric loss values at lower frequencies. Penetration depth tended to be greater when the direction of propagation was perpendicular to the muscle fiber.

Rapid and quantitative detection of the microbial spoilage in chicken meat by diffuse reflectance spectroscopy (600-1100 nm).

AIMS: To evaluate the feasibility of visible and short-wavelength near-infrared (SW-NIR) diffuse reflectance spectroscopy (600-1100 nm) to quantify the microbial loads in chicken meat and to develop a rapid methodology for monitoring the onset of spoilage. METHODS AND RESULTS: Twenty-four prepackaged fresh chicken breast muscle samples were prepared and stored at 21 degrees C for 24 h. Visible and SW-NIR was used to detect and quantify the microbial loads in chicken breast muscle at time intervals of 0, 2, 4, 6, 8, 10, 12 and 24 h. Spectra were collected in the diffuse reflectance mode (600-1100 nm). Total aerobic plate count (APC) of each sample was determined by the spread plate method at 32 degrees C for 48 h. Principal component analysis (PCA) and partial least squares (PLS) based prediction models were developed. PCA analysis showed clear segregation of samples held 8 h or longer compared with 0-h control. An optimum PLS model required eight latent variables for chicken muscle (R = 0.91, SEP = 0.48 log CFU g(-1)). CONCLUSIONS: Visible and SW-NIR combined with PCA is capable of perceiving the change of the microbial loads in chicken muscle once the APC increases slightly above 1 log cycle. Accurate quantification of the bacterial loads in chicken muscle can be calculated from the PLS-based prediction method. SIGNIFICANCE AND THE IMPACT OF THE STUDY: Visible and SW-NIR spectroscopy is a technique with a considerable potential for monitoring food safety and food spoilage. Visible and SW-NIR can acquire a metabolic snapshot and quantify the microbial loads of food samples rapidly, accurately, and noninvasively. This method would allow for more expeditious applications of quality control in food industries.

Detection of sodium chloride in cured salmon roe by SW-NIR spectroscopy.

Salt and moisture content of cured salmon roe (ikura) was determined using short-wavelength-near-infrared (SW-NIR) reflectance spectroscopy (600-1100 nm). SW-NIR can be used to measure chloride species. Calibrations for salt in bulk samples of high-quality roe (R(2) = 0.904, SEP = 0.421%, RPD = 3.21) and average-quality roe (R(2) = 0.711, SEP = 1.13%, RPD = 1.81), crushed eggs (R(2) = 0.857, SEP = 0.509%, RPD = 2.62), and individual eggs (R(2) = 0.731, SEP = 0.698%, RPD = 1.98) were developed using partial least squares (PLS) regression models. The heterogeneous distribution of lipid and moisture in the individual eggs limit the sensitivity of this method; however, this method provides a rapid nondestructive method for high-value food products where destructive testing is expensive or impractical and for process control applications.

Biochemical and functional properties of Atlantic salmon (Salmo salar) muscle proteins hydrolyzed with various alkaline proteases.

Protein hydrolysates (5, 10, and 15% degrees of hydrolysis) were made from minced salmon muscle treated with one of four alkaline proteases (Alcalase 2.4L, Flavourzyme 1000L, Corolase PN-L, and Corolase 7089) or endogenous digestive proteases. Reaction conditions were controlled at pH 7.5, 40 degrees C, and 7.5% protein content, and enzymes were added on the basis of standardized activity units (Azocoll units). Proteases were heat inactivated, insoluble and unhydrolyzed material was centrifuged out, and soluble protein fractions were recovered and lyophilized. Substrate specificities for the proteases was clearly different. Protein content for the hydrolysates ranged from 71.7 to 88.4%, and lipid content was very low. Nitrogen recovery ranged from 40.6 to 79.9%. The nitrogen solubility index was comparable to that of egg albumin and ranged from 92.4 to 99.7%. Solubility was high over a wide range of pH. The water-holding capacity of fish protein hydrolysates added at 1.5% in a model food system of frozen minced salmon patties was tested. Drip loss was on average lower for the fish protein hydrolysates than for egg albumin and soy protein concentrate, especially for Alcalase hydrolysates. Emulsification capacity for fish protein hydrolysates ranged quite a bit (75-299 mL of oil emulsified per 200 mg of protein), and some were better than soy protein concentrate (180 mL of oil emulsified per 200 mg of protein), but egg albumin had the highest emulsifying capacity (417 mL of oil emulsified per 200 mg of protein). Emulsification stability for fish protein hydrolysates (50-70%) was similar to or lower than those of egg albumin (73%) or soy protein concentrate (68%). Fat absorption was greater for 5 and 10% degrees of hydrolysis fish protein hydrolysates (3.22-5.90 mL of oil/g of protein) than for 15% hydrolysates, and all had greater fat absorption than egg albumin (2. 36 mL of oil/g of protein) or soy protein concentrate (2.90 mL of oil/g of protein).

Fish protein hydrolysates: production, biochemical, and functional properties.

Considerable amounts of fish processing byproducts are discarded each year. By developing enzyme technologies for protein recovery and modification, production of a broad spectrum of food ingredients and industrial products may be possible. Hydrolyzed vegetable and milk proteins are widely used food ingredients. There are few hydrolyzed fish protein foods with the exception of East Asian condiments and sauces. This review describes various manufacturing techniques for fish protein hydrolysates using acid, base, endogenous enzymes, and added bacterial or digestive proteases. The chemical and biochemical characteristics of hydrolyzed fish proteins are discussed. In addition, functional properties of fish protein hydrolysates are described, including solubility, water-holding capacity, emulsification, and foam-forming ability. Possible applications of fish protein hydrolysates in food systems are provided, and comparison with other food protein hydrolysates where pertinent.

Fatty acid composition of salmonid muscle changes in response to a high oleic acid diet.

Substitution of high oleic acid sunflower oil for herring oil in formulated salmonid diets affected the fatty acid composition of muscle, liver and visceral fat from coho salmon (Oncorhynchus kisutch) and rainbow trout (O. mykiss). Fish were fed diets containing either high oleic acid sunflower oil or herring oil as the supplemental lipid source (12.4 g/100 g diet) for 1-2 mo. Muscle from fish fed the sunflower oil diet had twice the concentration of oleic acid (approximately 25 g/100 g lipid) as muscle from fish fed the herring oil diet (approximately 12 g/100 g lipid). The maximum concentration of oleic acid in the muscle was obtained after only 2 wk of feeding the sunflower oil diet. Oleic acid concentrations in liver and visceral fat of fish fed the sunflower oil diet were significantly higher than in fish fed the herring oil diet. Rainbow trout fed the sunflower oil diet for 4 wk maintained the higher oleic acid concentrations in muscle and liver when deprived of feed for 2 wk compared with fish fed the herring oil diet. These data indicated that accumulation of oleic acid in coho salmon and rainbow trout muscle was fairly rapidly achieved when a high oleic acid diet was fed. The differences between the fish receiving the two dietary treatments in fatty acid composition and in concentrations of thiobarbituric acid-reactive substances in muscle stored at refrigerated temperatures were consistent with previously reported differences in aroma perceived by a sensory panel.

Irradiated or aseptically prepared frozen dairy desserts: acceptability to bone marrow transplant recipients.

Sterile ice cream and frozen yogurt were offered to immunosuppressed patients recovering from bone marrow transplantation. To obtain sterile products, two of the dairy desserts (prepackaged ice cream and frozen yogurt bars) were exposed to 40 kGy of cobalt 60 irradiation. Four different flavors of ice cream were aseptically prepared under a laminar airflow hood using commercially sterilized ingredients. A commercially sterile, frozen milk-based drink on the low-microbial menu served as the control. Ratings of the seven products by 17 patients indicated that a frozen vanilla milk-based drink and aseptically prepared chocolate ice cream were highly acceptable to recovery immunosuppressed patients who have difficulty eating most foods. However, the seven desserts received higher ratings from a sensory panel of healthy individuals than from the patient panel, confirming that new foods for the low-microbial diet should be "market-tested" by the targeted patient population before inclusion in the menu.

Application of immunochemical assays to food analysis.

Immunochemical assays are powerful bioanalytical techniques with application to several areas in food science, including food analysis, microbiology, nutrition, food safety, food quality, and process control. In principle, immunochemical techniques can be applied to the analysis of any compound, with only one specific antibody needed that can be obtained either from laboratory animals or, when available, from commercial sources. A well-designed immunochemical assay can detect targeted compounds at levels as low as 10(-12) M. Immunochemical techniques require little or no sample pretreatment, making these analytical procedures relatively rapid. The initial cost of developing an immunoanalytical assay may be high, but when the procedure is well established, the cost per test is often a fraction of that for other analytical methods. For these reasons, immunoanalytical assays provide an attractive alternative for the food analyst who requires either inexpensive qualitative screening tests or reliable quantitative methods with a high degree of sensitivity. This review concentrates on the use of enzyme immunoassay to address analytical problems in food chemistry and the analysis of various food components.

A comparison of dogfish and porcine pancreatic lipases.

1. A 16-fold purification of lipase activity from the pancreas of the dogfish (Squalus acanthias) was obtained by preparative isoelectric focussing. 2. The dogfish enzyme has a higher pH optimum (8.5) and a broader spectrum of activity above and below its optimum than a commercial porcine lipase preparation (optimum, 7.5). 3. The temperature optimum of dogfish lipase was 35 degrees C and it displayed greater stability than the porcine lipase preparation towards higher temperatures in the absence of substrate. 4. Dogfish lipase was active against a broad range of triacylglycerols and a wax ester. It had greater relative activity against the latter than the porcine lipase but hydrolyzed low mol. wt fatty acids less readily. In general, the effect of temperature was less on the dogfish enzyme than the porcine one.