Production of biogenic amines by<Emphasis Type="Italic"> Morganella morganii</Emphasis>,<Emphasis Type="Italic"> Klebsiella pneumoniae</Emphasis> and<Emphasis Type="Italic"> Hafnia alvei</Emphasis> using a rapid HPLC method

The histidine decarboxylating activity and production of biogenic amines by Morganella morganii (NCIMB, 10466), Klebsiella pneumoniae (NCIMB, 673) and Hafnia alvei (NCIMB, 11999) were investigated using a rapid HPLC method. Derivatisation of the bacterial samples was carried out using benzoyl chloride. A gradient elution system was used for analysis with a mixture of acetonitrile and HPLC grade water. Bacterial strains not only produce histamine in histidine-enriched broth but also the other biogenic amines. The chromatographic results show that bacterial strains are also capable of producing spermine and spermidine in histidine-enriched broth. Bacterial ammonia production by all three strains was clearly detected since ammonia is generated during the degradation of histidine. The study demonstrates that the highest histamine production was obtained by Morganella morganii, followed by Klebsiella pneumoniae, and the lowest with the Hafnia alvei. Therefore, Morganella morganii and Klebsiella pneumoniae have strong histidine decarboxylase activity since they are prolific histamine-forming bacteria     

Effects of process parameters on growth and metabolism of<Emphasis Type="Italic"> Lactobacillus sanfranciscensis</Emphasis> and<Emphasis Type="Italic"> Candida humilis</Emphasis> during rye sourdough fermentation

The effects of temperature, pH, inoculum level, and NaCl on the growth and metabolism of Lactobacillus sanfranciscensis and Candida humilis in rye sourdough were determined. The temperature optima for growth of C. humilis and L. sanfranciscensis were 28 and 32 °C, respectively. Yeast growth was inhibited at 35 °C. The pH did not affect yeast growth in the range 3.5–5.5, whereas growth of L. sanfranciscensis was inhibited at pH 4.0. A NaCl concentration of 4% (flour base) inhibited growth of L. sanfranciscensis but not C. humilis. The effects of the process parameters on the formation of lactate, acetate, ethanol, and CO₂ by the organisms were generally in agreement with their effects on growth. However, decreased formation of acetate by L. sanfranciscensis was observed at 35 °C although lactate and ethanol formation were not affected. In conclusion, the study provides a rationale for the stable persistence of L. sanfranciscensis and C. humilis in traditional sourdoughs and will facilitate the optimisation of sourdough fermentations in traditional and new applications.     

Formation of guaiacol from vanillin by<Emphasis Type="Italic"> Alicyclobacillus acidoterrestris</Emphasis> in apple juice: a model study

In this study, the formation of guaiacol from vanillin by Alicyclobacillus acidoterrestris in apple juice was investigated. Apple juices spiked with 10 and 100 mg/L of vanillin were inoculated with 1×10³ CFU/mL and 1×10⁵ CFU/mL spores of A. acidoterrestris and incubated at 25 and 46 °C. A. acidoterrestris started to form guaiacol from vanillin when the spore count exceeded the critical level of 1×10⁴ CFU/mL. Increasing the vanillin concentration also increased the concentration of guaiacol formed, especially at 46 °C. The growth of A. acidoterrestris was not favorable at 25 °C. Thus, the formation of guaiacol is strongly limited at room temperature.     

Formation of biogenic amines by Gram-negative rods isolated from fresh,spoiled, VP-packed and MAP-packed herring (<Emphasis Type="Italic">Clupea harengus</Emphasis>)

Bacterial strains (120) were isolated from fresh, spoiled, VP-packed and MAP-packed herring. Identified bacterial strains were investigated for their abilities to produce biogenic amines in histidine, lysine and ornithine decarboxylase broth by a rapid high-performance liquid chromatography (HPLC) method. The microflora of fresh herring was dominantly Pseudomonas (30%), Enterobacteriaceae (23.2%), Vibrio (13.3%) and Moraxella (13.3%) but, the microflora of herring stored in VP and MAP was dominated by species belonging to Vibrio (23.3%) and Moraxella (20%), which indicates that these packaging systems prevented the growth of Pseudomonas and Enterobacteriaceae. In a laboratory medium containing amino acid (histidine, ornithine and lysine), most of bacterial strains produced histamine, putrescine and cadaverine. The highest histidine decarboxylase activities were observed in Klebsiella oxytoca, Hafnia alvei and Proteus vulgaris which produced 396, 232 and 54 mg histamine/L, respectively in histidine-enriched broth. The accumulation of cadaverine by Klebsiella oxytoca and Hafnia alvei was 325 and 208 mg/l, respectively. All strains isolated produced putrescine in an ornithine-enriched broth, ranging from 3 to 249 mg/l. The production of putrescine by Klebsiella oxytoca and Hafnia alvei was 249 and 195 mg/l, respectively. Moraxella spp and Acinetobacter spp did not produce histamine which indicates they did not have histidine decarboxylase activity.     

Tyramine production of technological important strains of <Emphasis Type="Italic">Lactobacillus</Emphasis>, <Emphasis Type="Italic">Lactococcus</Emphasis> and <Emphasis Type="Italic">Streptococcus</Emphasis>

The aim of this paper was to study the biogenic amines (histamine, tyramine, putrescine, cadaverine, agmatine, spermine and spermidine) production of selected technological important lactic acid bacteria (strains of the genera Lactococcus, Lactobacillus and Streptococcus). Three methods (ion-exchange chromatography (IEC), PCR and cultivation method with pH indicator) were used. Within the 39 strains of lactic acid bacteria tested, the production of tyramine (formed by tyrosine decarboxylase) was detected in eight strains (3 strains of Lactococcus lactis subsp. lactis, three strains of Lactococcus lactis subsp. cremoris, 1 strain of Streptococcus thermophilus and 1 strain of Lactobacillus delbrueckii subsp. bulgaricus). The other tested biogenic amines were not detected. Cultivation in decarboxylation broth seems to be the least accurate method for the detection of biogenic amines due to enhanced risk of false-positive reactions. Therefore, in order to detect bacteria producing biogenic amines, the combination of PCR and chromatographic methods (e.g. IEC) can be recommended.     

The effects of different technologies on<Emphasis Type="Italic"> Alicyclobacillus acidoterrestris</Emphasis> during apple juice production

In this study, the effects of various processing steps applied during apple juice processing on Alicyclobacillus acidoterrestris were investigated. Raw apple juice was inoculated with A. acidoterrestris spores at two inoculum levels of 1×10³ cfu/ml and 1×10⁶ cfu/ml. Following enzymatic treatment, the raw juice was processed into clear juice using either conventional clarification or ultrafiltration. The number of A. acidoterrestris spores in the final product was determined to be dependent on both initial contamination level and processing conditions. Increasing temperature to 50 °C during depectinization resulted in a higher spore counts at both inoculum levels. Even if the ultrafiltration process was found to be much more convenient for the retention of A. acidoterrestris when compared to conventional clarification, the spores could penetrate the ultrafiltration membranes having both 20 and 50 kDa. Increasing membrane pore size and initial spore counts in the feed solution also increased the number of spores penetrating the membrane during ultrafiltration.     

Bacterial Growth and Histamine Production on Vacuum Packaged Tuna

Vacuum and nonvacuum packaged samples of tuna or tuna spiked with Klebsiella oxytoca T₂, Morganella morganii JM or Hafnia alvei T₈ were stored at 2°C and 10°C and examined for bacterial growth and histamine production on day 3, 6, 10 and 15. Unlike those stored at 2°C, tuna samples stored at 100°C showed significant bacterial growth over time and had high levels of histamine (>200 mg/100g tuna on day 15). Vacuum packaging did not show any beneficial effect in controlling bacterial growth and histamine production on tuna samples at these two temperatures. Low temperature storage was more effective than vacuum packaging in controlling histamine production by the bacteria tested.     

Adsorption isotherms of osmo-oven dried african star apple (<Emphasis Type="Italic">Chrysophyllum albidum</Emphasis>) and african mango (<Emphasis Type="Italic">Irvingia gabonensis</Emphasis>) slices

Halves of deseeded and deskinned pulps of African star apple (Chrysophyllum albidum) at three different stages of ripeness were immersed in sucrose solutions of 44°B, 52°B and 60°B in water baths at 27 °C and 40 °C for 8 h. About 10 mm slices of African mango (Irvingia gabonensis) at two different stages of ripeness were immersed in osmotic (sucrose) solutions of 52°B, 60°B and 68°B in water baths at 27 °C and 40 °C for 10 h. The adsorption isotherms of the osmo-oven dried fruit pieces were conducted at water activities between 0.1 and 0.9, at 20 °C and 40 °C. Higher equilibrium temperature and solids gained in the osmo-air dried pieces caused a depression of the adsorption isotherms, especially at low and intermediate a_w range. Crossing of the 20 °C and 40 °C isotherms for both African star apple and African mango occurred at a_w ~0.8. Sorption data was fitted into eight isotherm models. Guggenheim–Anderson de Boer (GAB) model was best fitted (r²=0.912 for African mango, and r²=0.998) for modelling and calculation of the monolayer moisture (M_m) content. M_m content increased with increasing degree of ripeness and total solids of osmo-oven dated products. M_m decreased with increased temperature. M_m of African mango and African star apple was in the range of 2.91–6.55 and 7.66–16.04% d.b., respectively.     

Prediction of<Emphasis Type="Italic"> Bacillus subtilis</Emphasis> spore survival after a combined non-isothermal-isothermal heat treatment

Different inactivation kinetics data have been used to predict the number of survivors exposed to a heat treatment and, in consequence, to design thermal processes for the food industry. In this work, spores of an acidophilic strain of Bacillus subtilis were heated under isothermal and non-isothermal conditions. Experimental results obtained after isothermal treatments were analysed using the classical two-step linear regression procedure and a one-step non-linear regression method. Data obtained after non-isothermal treatments were analysed using a one-step, non-linear procedure. Kinetic parameters obtained from isothermal heating were close, either using the two-step linear regression (D₁₀₀=6.5 min) or the one-step non-linear regression (D₁₀₀=6.3 min), although the second method gave smaller 95% confidence intervals. The z values derived from non-isothermal heating were higher than those obtained in isothermal conditions (z=9.3 °C for non-isothermal heating at 1 °C/min versus z=7.7 °C for isothermal heating one step non-linear regression). Results were validated with experimental data obtained after different heat treatments, consisting of a phase of temperature increase at a fixed rate, followed by a holding phase. Non-isothermal methods predicted accurately the number of survivors after the heating ramp, while isothermal methods were more accurate for the holding phase of the treatment. When a temperature profile of a typical heat treatment process applied in the food industry was simulated, all predictions were on the safe side.     

A New Multiplexed Real-Time PCR Assay to Detect <Emphasis Type="Italic">Campylobacter jejuni</Emphasis>, <Emphasis Type="Italic">C. coli</Emphasis>, <Emphasis Type="Italic">C. lari</Emphasis>, and <Emphasis Type="Italic">C. upsaliensis</Emphasis>

A new rapid method based on real-time PCR was developed to detect four thermophilic Campylobacter species (Campylobacter jejuni, Campylobacter coli, Campylobacter lari, and Campylobacter upsaliensis) in food samples. The assay targeted the bipA gene for C. upsaliensis and C. lari, whereas the gene encoding the ATP-binding protein CJE0832 was used to detect C. coli and C. jejuni. These genes were chosen for this assay due to their low variability and mutation rate at a species level. The multiplex PCR showed 100% inclusivity for all 25 thermophilic Campylobacter strains tested and 100% exclusivity for 38 non-targeted strains belonging to closely related species. The newly developed real-time PCR could detect down to 10² genomes/reaction and displayed efficiency above 97% for all species except for C. upsaliensis (90.1%). The method proved to be a reliable tool for food analysis, showing 100% sensitivity, 96% efficiency, and 92.45% specificity when validated against the gold standard method UNE-EN ISO 10272:2006 using 200 diverse food samples (meat, fish, fruits and vegetables, and raw milk). In artificially spiked samples, the detection limit of the method was 10 cfu/g in salad, 5 cfu/g in turkey meat, and 1 cfu/g in the rest of meat samples tested. Consequently, the newly designed molecular tool represents a quick and safe alternative to obtain reliable results concerning the presence/absence of the main thermophilic Campylobacter in any food sample.     

Thermal inactivation kinetics of quality-related enzymes in cauliflower (<Emphasis Type="Italic">Brassica oleracea</Emphasis> var. <Emphasis Type="Italic">botrytis</Emphasis><Emphasis Type="Bold">)</Emphasis>

Thermal inactivation of quality-related enzymes in both cauliflower crude enzyme extracts and fresh tissue samples was studied in temperature range 50–100 °C. For crude enzyme extracts, several parameters, reaction rate constants (k) and activation energy (E _a) as well as decimal reduction time (D) and (z) values, were used to characterize the thermal stability. The rates of inactivation were found to follow first-order inactivation kinetics. Activation energies varied between 101.18 and 208.42 kJ mol⁻¹ with z values of 10.59–24.09 °C. The examined kinetics indicated that lipoxygenase was the most heat resistant followed by peroxidase, polyphenol oxidase, pectin methyl esterase and ascorbic acid oxidase. Furthermore, the obtained results from the blanched fresh tissues indicated that inactivation of lipoxygenase secured disappearing of any other enzyme activities. Therefore, this study recommends using lipoxygenase as an indicator enzyme to optimize the thermal treatments of cauliflower products.     

Process optimisation of<Emphasis Type="Italic"> Gulab jamun</Emphasis>, an Indian traditional sweet,using sugar substitutes

Models capable of predicting the product quality of Gulab jamun using sugar and a sugar substitute have been developed using response surface methodology (RSM) and applied to determine the optimum processing conditions. Box–Behnken design was used considering three independent variables, namely concentration (°B) of syrup, its temperature and time of soaking, each at three different levels. The dependent variable or response measured for each treatment was a sensory score for overall quality, which is taken as a combination of the sensory impact of colour, appearance, texture, mouthfeel, taste and aroma. According to RSM, the optimum score for overall quality attained for Gulab jamun prepared with sugar was 7.7, obtained at processing conditions of 51°B syrup, 54 °C and 4 h soaking, whereas for the Gulab jamun prepared with the sugar substitute, sorbitol, a maximum overall quality score of 8.0 was obtained at processing conditions of 54°B syrup, 65 °C and 3 h soaking. These processing conditions were found to correlate with those of experimental conditions.     

Drying kinetics of whole and sliced <Emphasis Type="Italic">shiitake</Emphasis> mushrooms (<Emphasis Type="Italic">Lentinus edodes</Emphasis>)

Isotherms of shiitake mushroom (Lentinus edodes) at 25 and 40°C were determined and drying kinetics of whole and sliced shiitake mushrooms were tested using a convective air drying method at different drying temperature of 40, 50, 60, and 70°C. The monolayer moisture contents of the mushroom were 7.23 and 5.44 g water/100 g of dry solids at 25 and 40°C, respectively. Both mushroom samples showed falling drying rate periods with increasing drying rates with increases in drying temperature, and the drying rate of the sliced mushrooms was faster than that of the whole mushrooms at the same drying conditions. The kinetic parameters for dehydration of the mushrooms were determined using the Newton model and the Classical diffusion model. Activation energy (E _a ) values as determined using the Newton model were 22.58 and 20.48 kJ/mol for the whole and sliced mushrooms, respectively.     

Sourdough-isolated<Emphasis Type="Italic"> Lactobacillus fermentum</Emphasis> as a potent anti-mould preservative of a traditional Iranian bread

Sourdough bread is an ancient method for making long microbial shelf life and strongly flavoured breads. Traditional sourdough bread-making has been practised for centuries in Iran. Although rural bread-making is still highly reliant on sourdough, urban bakeries usually use bakers yeast and also sodium bicarbonate instead. In this study the anti-mould preservative effects of three lactic acid bacteria isolates were investigated on one of the popular traditional Iranian bread (Lavash). Starter cultures were prepared using 20 h cultures of three sourdough-isolated strains, Lactobacillus casei, Lactobacillus plantarum and Lactobacillus fermentum. Different concentrations (0.5%, 1%, 2%, 4% and 8%) of lactobacillus starter cultures were used to prepare a variety of sourdoughs. Lavash doughs were inoculated with 20% of each lactobacilli-fermented sourdough. After a 20 h incubation at 30 °C, a decrease in pH was observed in the different lactobacilli sourdoughs. However an 8% inoculum of L. fermentum resulted in a significant decrease in pH (3.87 to 2.70). Concentrations of 2% and higher of the three lactobacilli used in Lavash sourdough delayed mould growth during storage. However this preservative effect was more significant when an 8% inoculum of L. fermentum was used. These results tend to suggest that selected strains of lactobacilli have a beneficial role in prevention of bread waste by mould spoilage, and hence could extend bread shelf life.     

Influence of<Emphasis Type="Italic"> Lactobacillus plantarum</Emphasis> on<Emphasis Type="Italic"> Staphylococcus aureus</Emphasis> growth in a fresh vegetable model system

The influence of a Lactobacillus plantarum (B₄) on the growth of Staphylococcus aureus (Sa₄) was verified by impedometric methods in a suitable model reproducing the characteristics of fresh vegetables. The inoculum size of the single strains and their growth temperature were varied according to a Central Composite Design. The results obtained via statistical analysis showed that the temperature affected the growth of both S. aureus and L. plantarum strains. The pathogenic strain, independently of its inoculum size, was inhibited by L. plantarum at all the tested temperatures. A proper combination of specific lactic acid bacteria and storage temperature should improve the safety of the vegetable products.     

Influence of starter culture combinations of<Emphasis Type="Italic"> Lactobacillus fermentum</Emphasis>,<Emphasis Type="Italic"> Saccharomyces cerevisiae</Emphasis> and<Emphasis Type="Italic"> Candida krusei</Emphasis> on aroma in Ghanaian maize dough fermentation

Various combinations of Lactobacillus fermentum, Saccharomyces cerevisiae and Candida krusei were investigated as starter cultures for the fermentation of the Ghanaian maize dough. The four combinations used were each yeast species with L. fermentum, both yeast species together, and both yeast species together with L. fermentum. Maize dough fermented spontaneously was also used as control. The aroma profile, acidity values and sensory characteristics of all dough samples were compared. Using Principal Component Analysis (PCA), the aroma profiles of the dough samples could be separated into five distinct groups based on the type of fermentation. Fermentations where S. cerevisiae and C. krusei were combined produced the highest concentrations of aroma compounds, particularly esters, but were judged as having the lowest acceptability scores. A more balanced profile that included alcohols, carbonyls, esters and acids was obtained with the combination of L. fermentum and C. krusei, which produced fermented dough with a high sensory score comparable to spontaneous fermentations. Combinations of L. fermentum and S. cerevisiae produced high concentrations of ethanol and fusel alcohols. Lactic and acetic acid concentrations were highest with the combined use of all three starter cultures.     

Combined effect of thymol and cymene to control the growth of<Emphasis Type="Italic"> Bacillus cereus</Emphasis> vegetative cells

The antibacterial action of thymol and cymene on two Bacillus cereus strains (INRA-AVTZ415 and INRA-AVZ421) was studied. Thymol and cymene alone showed higher antibacterial effect on the growth of B. cereus in BHI at 30 °C, both in lag and in exponential growth phases, with increasing added concentrations. However, the two strains tested showed different sensitivity to these natural compounds. When thymol and cymene were applied simultaneously, it resulted in a greater antibacterial effect than when the compounds were applied separately. The synergistic behaviour of both compounds was also shown for lag phase B. cereus INRA-AVZ421 cells in food substrates (carrot juice, chicken soup and pumpkin cream) at 30 °C, though a clear food-related effect was observed. This study indicates the potential use of thymol and cymene applied simultaneously for preservation of minimally processed foods.     

Effect of soybean substitution for cowpea on physical,compositional, sensory and sorption properties of<Emphasis Type="Italic"> akara</Emphasis> Ogbomoso

Akara Ogbomoso was prepared from cowpea:soybean flour mixture ratios of 100:0, 80:20, 60:40 and 40:60. Physical, compositional, sensory and sorption characteristics of the akara were evaluated. Protein and fat content increased, while carbohydrate content decreased as the soy content of the flour mixture of akara Ogbomoso increased. There were no significant differences (P<0.01) in the colour, taste, aroma and overall acceptability for either the freshly prepared akara Ogbomoso or the akara Ogbomoso stored for 10 weeks. There were significant differences (P<0.01) in crunchiness, increasing as the soy substitution increased both in the freshly prepared and stored akara Ogbomoso. The adsorption isotherms of akara Ogbomoso varied with varying levels of soy substitution and at 20, 30 and 40 °C, had sigmoid shaped, type II isotherms according to Brunauer-Emmet-Teller (BET) classification, and were affected by temperature and composition. Increased levels of soy substitution and temperature depressed the isotherms. Calculated monolayer moisture contents using BET and Guggenhein Anderson deBoer models decreased with increasing soy substitution and temperature.     

Non-structural carbohydrates in processed soft fried onion (<Emphasis Type="Italic">Allium cepa</Emphasis> L.)

The effects of curing, removal of top and root, peeling, storage of raw and peeled onion on the content of non-structural carbohydrates and the ability of onion to develop a brown colour were studied. Glucose, fructose, sucrose, 1-kestose, neokestose, nystose, 1^F--d-fructofuranosylnystose and four unknown oligofructosides were quantified using high performance anion exchange chromatography coupled with pulsed amperometric detection (HPAEC-PAD). Field drying affected the concentration of fructose only. Glucose, fructose and an unknown fructan decreased in concentration while the concentration of, e.g. 1-kestose, neokestose and nystose increased by removal of root and top of the onions when stored at 15 °C for one week. Peeling had minor influence on changes in non-structural carbohydrates. Storage of non-peeled (raw) onions at 5 °C and storage of mechanically damaged and peeled onion at 20 °C promoted fructan hydrolysis and browning of soft fried onion. Similar changes were produced by long-term storage of cultivars of common onion. Dry matter and fructose increased during storage of peeled onions at both 5 and 20 °C. The increase in fructose content during storage was due to a significantly decrease in the content of 1-kestose, neokestose, 1^F--d-fructofuranosylnystose, an unknown fructan and total fructans. It was concluded that the missing ability of industrially processed onions to develop a brown colour may be overcome by storage of the peeled onions for a few hours or even overnight at 5 °C.