Use of immunological techniques for detecting species substitution in raw and smoked fish

 Immunological techniques based on double immunodiffusion and immunodotting have been designed to detect the substitution of halibut (Hippoglossus hippoglossus) for sole (Solea solea) fillets. An immunodotting technique has been also developed to differentiate between smoked cod (Gadus morhua) and smoked eel (Anguilla anguilla) fillets. Antisera raised against water-soluble extracts of raw halibut and smoked cod were employed. Antiserum to cod proteins did not show cross-reaction with eel proteins, whereas antiserum to halibut proteins cross-reacted with sole proteins. Species-specific antiserum to halibut proteins was achieved by adsorption on a polymer of sole proteins. These methods are very easy to perform and provide the basis for the development of rapid tests for detecting species substitution in fish products. Received: 18 April 1996/Revised version: 5 July 1996     

Human Milk Banking: Influence of Different Pasteurization Temperatures on Levels of Protein Sulfur Amino Acids and Some Free Amino Acids

Human milk is frequently heat‐treated in hospitals, particularly milk that is banked, to destroy contaminating bacteria and viruses, but this treatment simultaneously reduces the content of some vitamins, enzymes, and immunological and nutritional factors. This study was performed to find the optimal conditions for heat treatment. The effects of 2 pasteurization temperatures on levels of protein sulfur amino acids (methionine, cystine) and some free amino acids (taurine, glutamine, glutamic acid) in light of the oxidative instability that occurs especially during thermal treatment were examined. These substances in raw human milk and in milk treated at 56.5 °C and 62.5 °C for 30 min were compared. Samples of mature human milk from all feeds over 24 h were obtained from 13 healthy well‐nourished mothers of term infants. Each sample was divided into 3 parts: raw, treated at 56.5 °C for 30 min, treated at 62.5 °C for 30 min. The results showed that the availability of sulfur amino acids and free taurine is the same after heat treatment, whereas milk processing increased positively the levels of free glutamic acid and glutamine, but there is significance only for glutamine. The mean quantities of considered amino acids were similar in milk treated at the recommended pasteurization temperature (30 min at 62.5 °C) and at 56.5 ° for 30 min.     

Effect of Raw Potato Composition on Acrylamide Formation in Potato Chips

ABSTRACT: Recent studies have shown that subjecting foods to high temperatures during cooking processes such as frying gives rise to the formation of acrylamide. Several factors including product composition and processing conditions affect the rate of formation of this chemical in starch-rich foods. Low reducing sugar and the amino acid asparagine content is desired when cooking because the formation of acrylamide is attributed to the Maillard reaction that occurs between these food components. The cultivar 'Atlantic' was used to determine the effect of potato components (reducing sugars and asparagine) on acrylamide content during frying in a traditional fryer. A model system was developed by infusing leached potato slices with predetermined amounts of glucose and asparagine. Increasing glucose and asparagine content in the slices increased the acrylamide content in the potato chips. Color could not be used as an indication of acrylamide content because potato chips with similar color had very different acrylamide concentrations.     

The effect of red pigment on the amyloidization of yeast proteins

The intensity of amyloid-bound thioflavine T fluorescence was studied in crude lysates of yeast strains carrying mutations in the ADE1 or ADE2 genes and accumulating the red pigment (a result of polymerization of aminoimidazoleribotide), and in white isogenic strains--either adenine prototrophs or carrying mutations at the first stages of purine biosynthesis. We found that the red pigment leads to a drop of amyloid content. This result, along with the data on separation of protein polymers of white and red strains in PAGE, suggests that the red pigment inhibits amyloid fibril formation. The differences in transmission of the thioflavine T fluorescence pattern by cytoduction and in blot-hybridization of pellet proteins of red and white [PSI(+) ] strains with Sup35p antibodies confirmed this conclusion. Purified red pigment treatment also led to a decrease of fluorescence intensity of thioflavine T bound to insulin fibrils and to yeast pellet protein aggregates from [PSI(+) ] strains. This suggests red pigment interaction with amyloid fibrils. Comparison of pellet proteins from red and white isogenic strains separated by 2D-electrophoresis followed by MALDI analysis has allowed us to identify 48 pigment-dependent proteins. These proteins mostly belong to functional classes of chaperones and proteins involved in glucose metabolism, closely corresponding to prion-dependent proteins that we characterized previously. Also present were some proteins involved in stress response and proteolysis. We suppose that the red pigment acts by blocking certain sites on amyloid fibrils that, in some cases, can lead in vivo to interfere with their contacts with chaperones and the generation of prion seeds.     

Temperature effects on the removal of potential HFC replacements, CF3CH2CH2OH and CF3(CH2)2CH2OH, initiated by OH radicals

The gas-phase kinetic coefficients of OH radicals with two primary fluorinated alcohols, CF(3)CH(2)CH(2)OH (k(1)) and CF(3)(CH(2))(2)CH(2)OH (k(2)), potential replacements of hydrofluorocarbons (HFCs), are reported here as a function of temperature (T = 263-358 K) for the first time. k(1) and k(2) (together referred as k(i)) were measured under pseudo-first-order conditions with respect to the initial OH concentration using the pulsed laser photolysis/laser induced fluorescence technique. The observed temperature dependence of k(i) (in cm(3) molecule(-1) s(-1)) is described by the following Arrhenius expressions: k(1)(T) = (2.82 ± 1.28) × 10(-12) exp{-(302 ± 139)/T} cm(3) molecule(-1) s(-1) and k(2)(T) = (1.20 ± 0.73) × 10(-11) exp{-(425 ± 188)/T} cm(3) molecule(-1) s(-1).The uncertainties in the Arrhenius parameters are at a 95% confidence level (± 2σ). Uncertainties in k(i)(T) include both statistical and systematic errors. Activation energies were (2.5 ± 1.2) kJ/mol and (3.6 ± 1.6) kJ/mol for the OH-reaction with CF(3)CH(2)CH(2)OH and CF(3)(CH(2))(2)CH(2)OH, respectively. The global lifetime (τ) at 275 K for CF(3)CH(2)CH(2)OH and CF(3)(CH(2))(2)CH(2)OH due to the OH-reaction was estimated to be ca. 2 weeks and 5 days, respectively. The reported Arrhenius parameters can be used in 3D models that take into account the geographical region and season of emissions for estimating a matrix of instantaneous lifetimes. As a consequence of the substitution of the -CH(3) group by a -CH(2)OH group in HFCs, such as CF(3)CH(2)CH(3) and CF(3)(CH(2))(2)CH(3), the tropospheric lifetime with respect to the OH reaction is significantly shorter and, since their radiative forcing is similar, global warming potentials of CF(3)CH(2)CH(2)OH and CF(3)(CH(2))(2)CH(2)OH are negligible. Therefore, CF(3)CH(2)CH(2)OH and CF(3)(CH(2))(2)CH(2)OH seem to be suitable alternatives to HFCs.     

Polychlorinated dibenzo-p-dioxins and dibenzofurans in the air of Seveso,Italy, 26 years after the explosion

This study reports the current levels of polychlorinated dibenzo-p-dioxins (PCDDs) and furans (PCDFs) in air at Seveso, where an explosion in a 2,4,5,-trichlorophenol production reactor occurred 26 years ago. The aims were to assess if residues of the 2,3,7,8-tetrachlorodibenzo-p-dioxin (2,3,7,8-TCDD) released during the accident and still present in soil could contaminate the above air and to investigate other potential sources in the area. Long-term air collection was carried out in zones A and B in Seveso and in a reference location in Milan, and samples were analyzed for PCDD and PCDF concentrations by gas chromatography-mass spectrometry (GC-MS). Experimental results showed that no important contribution to the air concentrations is due to the soil contamination and that contemporary sources essentially control the atmospheric burden of PCDDs and PCDFs in the Seveso area. The theoretical release of 2,3,7,8-TCDD from the soils of zones A and B of Seveso was calculated using the SoilFug model. In the worst case, the model simulated an enrichment in atmospheric 2,3,7,8-TCDD concentrations of 4 and 22% for zones A and B, respectively. The investigation of the potential emission sources in the area indicated that combustion of wood residues from furniture factories may be an additional local source of PCDDs and PCDFs.     

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.     

Comparison of the stability of palm olein and a palm olein/canola oil blend during deep‐fat frying of chicken nuggets and French fries

Stability of palm olein (PO) and a blend 50% palm olein/50% canola oil (POC) during deep‐fat frying at 180 °C of French fries (FF) or chicken nuggets (CN) was studied through the determination of physical and chemical parameters in the fresh and used oils. Degradation at the end of the study resulted in total polar compounds of 12–13.5% for PO and 11.5–14.5% for POC and viscosity of 65–123.3 cP for PO and 63–72.8 cP for POC. Lower peroxide values (5.33–6.32) were obtained for the blend (PO had 5.21–8.55). Food type affected colour parameters and p‐anisidine value of the oils. For CN, the lowest fat content and higher hardness were obtained when they were fried in PO. CN caused a faster deterioration in the oils, in comparison with FF, especially in POC. Gas chromatography allowed to observe differences in fatty acids composition for both used oils.     

Polymerase chain reaction coupled with peptide nucleic acid high-performance liquid chromatography for the sensitive detection of traces of potentially allergenic hazelnut in foodstuffs

A new DNA extraction method suitable for a wide variety of complex food matrices has been devised and applied in combination with a new polymerase chain reaction (PCR) method for the sensitive detection of a specific DNA sequence univocally identifying the presence of potentially allergenic hazelnut (Corylus avellana). A 156 base pair amplicon corresponding to an internal region of the complementary DNA of the major hazelnut allergen (Cor a 1) was designed, and was found to be highly specific; the method was tested on both pure and complex food matrices and was found to be able to confirm the presence of hazelnut down to 5 pg of its DNA. The sequence of the amplicon was further confirmed by high-performance liquid chromatography (HPLC) analysis with a specific peptide nucleic acid (PNA) probe. A 15-mer PNA probe was expressly designed and synthesized to hybridize an internal sequence of the previously described amplicon. Given its reported sequence specificity and high hybridization efficiency, the PNA probe was used to develop an anion-exchange HPLC method allowing for a fast and reliable confirmation of the identity of the amplified products. The PCR–HPLC method was successfully tested on commercial samples, allowing for the detection of the presence of potentially hidden allergens even in products where the presence of hazelnut as an ingredient or possible contaminant was not reported.