Artur X. Roig-Sagués

Halotolerant and halophilic histamine-forming bacteria isolated during the ripening of salted anchovies (Engraulis encrasicholus).

This study was performed to investigate halotolerant and halophilic histamine-producing bacteria isolated during the ripening of salted anchovies. Of the isolates obtained during the ripening of anchovies, 1.37% showed histamine-forming activity, most of them (70%) belonging to the Staphylococcus genus. S. epidermidis showed a powerful histamine-forming activity, producing more than 1,000 microg/ml in the presence of 3% and 10% NaCl. Another powerful histamine-producing bacterium isolated during the ripening of salted anchovies was S. capitis. It was able to produce about 400 microg/ml of histamine in 10% NaCl under experimental conditions. Most of these species might be expected to be found as a result of contamination of fish during capture and subsequent unhygienic handling. However, no increase in histamine content was found in any batches through the ripening process. Histamine content always was acceptable in accordance with the maximum allowable levels of histamine fixed by the Spanish and European Union regulations.

Sensory quality and histamine formation during controlled decomposition of tuna (Thunnus thynnus)

Histamine production was studied during controlled tunafish decomposition at 0, 8, and 20°C. The influence of the location of the anatomic section on the amount of histamine formed and the incidence of histidine decarboxylating bacteria were also considered. By the time of sensory rejection, histamine levels in tunafish sections stored at 0 and 20°C were still below the hazard levels and the allowable levels established by both the U.S. Food and Drug Administration (FDA) and the European Union. Toxic amounts were only formed after the tunafish was considered organoleptically unsuitable for human consumption. However, at 8°C, levels of histamine between 100 and 200 mg/l00 g of fish were found before tuna reached the rejection point. Hence, physical appearance was not a good criterion for estimating the shelf life and especially the histamine-related health hazard when tuna was stored at 8°C, a common temperature in many home refrigerators.

Bacteriological Quality of Tuna Fish (Thunnus thynnus) Destined for Canning: Effect of Tuna Handling on Presence of Histidine Decarboxylase Bacteria and Histamine Level

The present paper analyzes the bacteriological quality and histamine content of tuna fish samples destined for canning. Raw material used in the canning process was of good quality. Histamine-producing bacteria counts were only found in three samples from the last step of the canning process before sterilization. Most of the bacteria identified as histamine formers were gram negative, and nearly all of which belonged to the Enterobacteriaceae family. Morganella morganii was the most frequent and active histamine former in tuna fish destined for canning. Other powerful histamine-producing bacteria isolated during the canning operation were Klebsiella oxytoca , Klebsiella pneumoniae and some strains of Enterobacter cloacae and Enterobacter aerogenes . They all were able to produce more than 500 ppm of histamine in experimental conditions. Most of these species might be expected to be found as a result of contamination of fish during capture and subsequent unhygienic handling in the canning plant. An increase in histamine content in tuna meat was not expected through the canning process. Histamine content always was acceptable in accordance with the maximum allowable levels of histamine fixed by both the European Economic Community and Food and Drug Administration.

Influence of ultra-high pressure homogenisation on antioxidant capacity, polyphenol and vitamin content of clear apple juice

Ultra-high pressure homogenisation (UHPH) is a recently developed technology and is still under study to evaluate its effect on different aspects of its application to food products. The aim of this research work was to evaluate the effect of UHPH treatments on quality characteristics of apple juice such as antioxidant capacity, polyphenol composition, vitamin C and provitamin A contents, in comparison with raw (R) and pasteurised (PA) apple juice. Several UHPH treatments that include combinations of pressure (100, 200 and 300MPa) and inlet temperatures (4 and 20°C) were assayed. Apple juice was pasteurised at 90°C for 4min. Antioxidant capacity was analysed using the oxygen radical antioxidant capacity (ORAC), 2,2-diphenyl-1-picrylhydrazyl (DPPH), trolox equivalent antioxidant capacity (TEAC), ferric reducing antioxidant power (FRAP) assay while total phenolic content was determined by the Folin-Ciocalteau assay. According to the FRAP and DPPH assays, UHPH processing did not change apple juice antioxidant capacity. However, significant differences were detected between samples analysed by TEAC and ORAC assays. In spite of these differences, high correlation values were found between the four antioxidant capacity assays, and also with total polyphenol content. The analysis and quantification of individual phenols by HPLC/DAD analytical technique reflects that UHPH-treatment prevented degradation of these compounds. Vitamin C concentrations did not change in UHPH treated samples, retaining the same value as in raw juice. However, significant losses were observed for provitamin A content, but lower than in PA samples. UHPH-treatments at 300MPa can be an alternative to thermal treatment in order to preserve apple juice quality.

Electrochemical detection of Salmonella using gold nanoparticles.

A disposable immunosensor for Salmonella enterica subsp. enterica serovar Typhimurium LT2 (S) detection using a magneto-immunoassay and gold nanoparticles (AuNPs) as label for electrochemical detection is developed. The immunosensor is based on the use of a screen-printed carbon electrode (SPCE) that incorporates a permanent magnet underneath. Salmonella containing samples (i.e. skimmed milk) have been tested by using anti-Salmonella magnetic beads (MBs-pSAb) as capture phase and sandwiching afterwards with AuNPs modified antibodies (sSAb-AuNPs) detected using differential pulse voltammetry (DPV). A detection limit of 143 cells mL(-1) and a linear range from 10(3) to 10(6) cells mL(-1) of Salmonella was obtained, with a coefficient of variation of about 2.4%. Recoveries of the sensor by spiking skimmed milk with different quantities of Salmonella of about 83% and 94% for 1.5×10(3) and 1.5×10(5) cells mL(-1) were obtained, respectively. This AuNPs detection technology combined with magnetic field application reports a limit of detection lower than the conventional commercial method carried out for comparison purposes in skimmed milk samples.

Inactivation of Listeria innocua in milk and orange juice by ultrahigh-pressure homogenization.

The aim of this work was to evaluate the bactericidal efficacy of ultrahigh-pressure homogenization (UHPH) against Listeria innocua ATCC 33090 inoculated into milk and orange juice. We also intended to study the effect of inlet temperature on the lethality and production of sublethal injuries in this microorganism and its ability to survive, repair, and grow in refrigerated storage after UHPH treatment. Samples of ultrahigh-temperature whole milk and ultrahigh-temperature orange juice inoculated at a concentration of approximately 7.0 log (CFU per milliliter) were immediately pressurized at 300 MPa on the primary homogenizing valve and at 30 MPa on the secondary valve, with inlet temperatures of 6.0 +/- 1.0 degrees C and 20 +/- 1.0 degrees C. L. innocua viable counts and injured cells were measured 2 h after UHPH treatment and after 3, 6, and 9 days of storage at 4 degrees C for milk and after 3, 6, 9, 12, 15, 18, and 21 days of storage at 4 degrees C for orange juice. Both the inlet temperature and the food matrix influenced significantly (P < 0.05) the inactivation of L. innocua, which was higher in whole milk at the 20 degrees C inlet temperature. The UHPH treatment caused few or no sublethal injuries in L. innocua. During storage at 4 degrees C after treatments, counts increased by approximately 2 logarithmic units from day 0 to 9 in whole milk, whereas in orange juice counts diminished by approximately 2.5 logarithmic units from day 0 to 18.

Inactivation by Ultrahigh-Pressure Homogenization of Escherichia coli Strains Inoculated into Orange Juice

The aim of this work was to evaluate the efficacy of ultrahigh-pressure homogenization (UHPH) for inactivation and/or sublethal injury of two strains of Escherichia coli (O58:H21 ATCC 10536 and O157:H7 CCUG 44857) inoculated into orange juice (pH 3.6). The effects of orange juice inlet temperature (6 and 20 degrees C) on the lethality values and the capacity of these strains for survival, repair, and growth during refrigerated storage after UHPH treatment also was evaluated. Samples of orange juice that had been treated with ultrahigh temperatures were inoculated with E. coli in the stationary phase of growth until a final concentration of approximately 7.0 log CFU/ml was reached. These samples were then treated for one cycle with a double-valve UHPH machine, with 300 MPa at the primary homogenizing valve and 30 MPa at the secondary valve. Counts of viable and injured bacterial cells were obtained for samples taken 2 h after UHPH treatment and after 3, 6, 9, 12, 15, 18, 21, 27, and 33 days of storage at 4 degrees C. The inlet temperature and the strain type both influenced significantly (P < 0.05) the lethality effect on E. coli, which was higher when the inlet temperature was 20 degrees C. No sublethal injuries were detected after any treatment. The changes in viable counts over time for both strains in pressurized and control samples were similar. The viable counts remained high from day 0 to day 18 and then tended to decrease. After 27 days of storage at 4 degrees C, E. coli O157: H7 was more resistant in orange juice samples pressurized at inlet temperatures of 6 and 20 degrees C, with viable counts of 3.41 and 3.20 log CFU/ml, respectively.

Evaluation of biogenic amines and microbial counts throughout the ripening of goat cheeses from pasteurized and raw milk

The effect of the hygienic quality of milk on changes in microbial counts and biogenic amine content was evaluated during ripening of goat cheeses manufactured from pasteurized and raw milks at 1, 14, 30, 60 and 90 d. The original milk, rennet, curd and whey were also included in the study. The pH, salt content and extent of proteolysis in the cheese were also evaluated. Spermidine and spermine were the main amines in raw milk, while they were minor amines in cheeses. Other amines increased markedly during ripening, tyramine being the main amine in cheese made from raw milk and cadaverine and putrescine in those produced from pasteurized milk. Enterobacteriaceae counts decreased during ripening whereas those of lactic acid bacteria increased, especially lactobacilli and enterococci. Cheese made from raw milk showed higher microbial counts during ripening than those made from pasteurized milk, especially for Enterobacteriaceae and enterococci, counts being 2 or 3 log units higher. Raw milk cheese showed remarkably higher biogenic amines compared with pasteurized milk cheeses. Therefore, pasteurization of milk causes a decrease in final biogenic amine content of cheese as a result of the reduction of its microbial counts.

Histidine decarboxylase activity of bacteria isolated from raw and ripened Salchichón, a Spanich cured sausage

Twenty samples of raw sausages (before the ripening process) and 10 of ripened sausages of two different sizes of salchichón , one of the most-consumed ripened meat products in Spain, were microbiologically analyzed. Histidine decarboxylase activity of different isolates obtained from different culture media was evaluated by a decarboxylase agar medium and confirmed afterwards by an enzymatic method. Of the isolates obtained from raw sausages, 15.8% showed histamine-forming activity, most of them (78.1%) belonging to the Enterobacteriaceae family. Klebsiella oxytoca , Enterobacter aerogenes (one isolate), and Enterobacter cloacae (six isolates) showed a powerful histamine-forming activity, producing more than 3,000 μg/ml. In spite of histamine-forming activity being detected in eight of ten analyzed samples of ripened sausages, only one histamine-forming bacterium, identified as Lactobacillus curvatus , was isolated; it showed an ability to form 1,994 μg/ml of histamine in experimental conditions.

Use of ultra-high-pressure homogenization to preserve apple juice without heat damage

Ultra-high-pressure homogenization (UHPH) is a technology for continuous treatment of fluid food products and can be applied to avoid thermal treatment of fruit juices. Treatment of fresh apple juice at homogenization pressures from 100 to 300 MPa and inlet temperatures of 4 °C or 20 °C caused important decreases in microbial counts when treatment pressure reached at least 200 MPa. After the treatment, counts were<1 log cfu/ml when homogenization pressure reached at least 200 MPa, and remained stable for more than 2 months at 4 °C. Hydroxymethylfurfural for thermal-treated juice reached values 100-fold higher than for its UHPH-treated or raw counterparts. Browning index was higher in non-thermal-treated juice, and it had an inverse correlation with the severity of UHPH treatment. Part of this browning occurred during Pilot Plant processing, but it kept on browning during preservation in juices in which PPO was not fully inactivated. UHPH treatment of apple juice may be an alternative to conventional thermal processing.