An evaluation of the effect of storage and processing temperatures on the microbiological status of sous vide extended shelf-life products

The microbiological and organoleptic quality of selected sous vide products was monitored over a storage period of up to five weeks at 3°C and 8°C, respectively. Sous vide products stored at the recommended temperature of 3°C showed negligible microbial growth and were found to be organoleptically acceptable throughout the storage period. Total plate counts at the end of the fourth week of storage were in the range <10–7×10³ CFU/g for all 19 products under study. Listeria monocytogenes, Salmonella, Clostridium perfringens, Bacillus cereus and Enterobacteriaceae were not detected in any of the processed samples. At 8°C, under temperature abuse conditions, while some products had acceptable microbial levels of 10²–10⁴ CFU/g after three weeks, others such as chicken chasseur had counts above 10⁶ CFU/g by the second week of storage. Lactic acid bacteria and Pseudomonas species were dominant in the microbial flora of spoilt samples and B. cereus (>3×10⁴ CFU/g) was isolated from spoilt chicken chasseur samples in the fourth week of storage at 8°C. Critical factors affecting sous vide product safety are discussed.     

Survival and growth of Listeria monocytogenes on sausage formulated with inoculated and stored rework product

To assess the effect of including contaminated rework on survival and growth of Listeria monocytogenes, two sausage formulations (one American, Bologna sausage; and one Bulgarian, Stranja sausage) were inoculated with the pathogen and stored for 4 days at 10 °C plus 15 h at 30 °C. After storage, both rework types were included (at 20% and 40%) in corresponding fresh sausage emulsions and heated to 68, 70 and 71.7 °C; fresh Bologna and Stranja emulsions served as controls and were inoculated with 24 h broth cultures of the same 10-strain mixture of L. monocytogenes and thermally treated to the same temperatures. The results showed that heating to 68 and 70 °C inactivated 3–4 log CFU/g of the initial concentration of L. monocytogenes cells (>7 log CFU/g), while heat treatment to 71.7 °C in the center of experimental samples reduced counts by 6 log CFU/g. Survival of L. monocytogenes in samples heated to 68 and 70 °C was higher in controls. Control samples of Stranja emulsion heated to 71.7 °C allowed higher growth (P < 0.05) during storage (5 days at 10 °C) as compared to other control and experimental rework samples. The Stranja emulsion had a higher fat content (20.2%) compared to the Bologna emulsion (11%). This study provides evidence about the possible danger when potentially contaminated rework is stored and then introduced into fresh product formulations.     

Characteristics of Bacillus cereus isolates from legume-based Indian fermented foods

An antibiogram of 48 strains of Bacillus cereus isolated from 6 different kinds of legume-based Indian fermented foods (amriti, dhokla, dosa, idli, papad and wadi) was generated against 18 different antibiotics that are commonly used against foodborne diseases, mainly gastroenteritis. Each of the isolates was found to be resistant against at least nine different antibiotics. Production of extracellular enzymes, namely protease, lipase and amylase by 33%, 27% and 46%, respectively, of the isolates indicates their potentiality for food spoilage. In brain–heart infusion broth supplemented with glucose, the D_100 °C-values for the tested 12 strains ranged from 3.0 to 9.2 min. In nutrient broth, the minimum and maximum pHs permitting growth of B. cereus were 5.3 and 11.6, respectively. The minimum inhibitory concentrations of sodium chloride, benzoic acid and sorbic acid for the growth of the isolates were 65–85 mg ml⁻¹, 400–700 μg ml⁻¹ (pH 5.0–4.2) and 500–600 μg ml⁻¹ (pH 5.0–4.8), respectively. Of the tested 10 strains, eight were resistant to 300 μg nisin ml⁻¹ (pH 5.0). While studying the combined effect of selected hurdles on the growth of an isolate, the judicious combination considered was 20 mg sodium chloride, 300 μg benzoic acid and 25 μg nisin ml⁻¹ at pH 5.6. The whole-cell protein fingerprinting (WCPF) analysis using SDS–PAGE revealed a high level of diversity among the isolates. At 60% similarity level, the WCPF profiles could be grouped into four major clusters which were divided into 34 subclusters. Most of the subclusters were source-wise homogeneous.     

Antimicrobial efficacy of cinnamon oil against Salmonella in almond based matrices

Reduced moisture enhances resistance of Salmonella and subsequently reduces the antimicrobial efficacy of thermal treatment. Alternative and supplementary non-thermal intervention methods are urgently needed. In this study, Cinnamonum cassia oil was tested for its antimicrobial effect against outbreak strains Salmonella Enteritidis PT30 and S. Tennessee K4643. Minimal inhibitory concentration and minimal bactericidal concentration for both strains were 0.05% (v/v) and 0.1% (v/v), respectively. Death curves showed that including 0.1% and 0.15% (v/v) C. cassia oil resulted in ∼7 Log reduction of bacteria within 2 h and 1 h, respectively. However, the antimicrobial efficacy of C. cassia oil was reduced when S. Enteritidis PT30 existed in low moisture condition. When S. Enteritidis PT30 was established on almonds/paper discs, 0.4% C. cassia oil resulted in ∼1.7 Log₁₀ CFU/almond or 3.2 Log₁₀ CFU/disc reduction within 2 h at room temperature, respectively. S. Enteritidis PT30 established on both almonds and paper discs were very stable, there was only a 0.80 Log₁₀ CFU/almond and 1.20 Log₁₀ CFU/disc reduction during 9-week and 7-week storage at room temperature, respectively. C. cassia oil intervention increased S. Enteritidis PT30 reduction on both almonds and paper discs during storage with more reduction on paper discs. 0.4% C. cassia oil treatment reduced S. Enteritidis PT30 on paper disc to undetectable level within 4 weeks, but only led to 2 Log₁₀ CFU reduction on almonds, indicating a protection effect from the almond matrix or almond surface components. Additionally, S. Enteritidis PT30 established on paper disc coated with almond surface components exhibited higher resistance to desiccation and C. cassia oil treatment, further demonstrating the protection role of food matrix. In conclusion, C. cassia oil is effective against S. Enteritidis PT30 and S. Tennessee K4643, but its antimicrobial efficacy against the tested Salmonella was compromised by low moisture environment and food matrix.     

Microbiological profile of helva sold at retail markets in Izmir city and the survival of Staphylococcus aureus in this product

The objective of this research was to determine the microbiological profile of tahin helva, a low moisture confectionery. A total of 63 tahin helva samples were collected from the retail markets in Izmir, Turkey. These samples were examined by standard procedures for aerobic plate count (APC) and counts of moulds and yeasts, Enterobacteriaceae, Staphylococcus aureus and Salmonella spp. APC of samples ranged from <10 to 1.6 × 10⁵ cfu/g. The counts for moulds, yeasts and Enterobacteriaceae ranged from <10 to 1.8 × 10³, <10 to 7 × 10², <10 to 8.5 × 10² cfu/g, respectively. None of the samples contained S. aureus or Salmonella. The potential for survival of S. aureus in the product stored at refrigeration (4 °C) and room (20 °C) temperatures was evaluated by artificial contamination. S. aureus cells were still recovered after 9 months of storage at both temperatures. The results of this investigation indicate that S. aureus contamination of tahin helva, (a_w of 0.172) may constitute a potential public health hazard, depending on the extent of contamination and mishandled exercise at the sale area and the kitchen.     

Reducing nitrite content in hot dogs by hurdle technology

The objective of the present study was to reduce nitrite content in hot dogs using hurdle technology without sacrificing product safety and quality. In the present study, the water activity of the hot dog was adjusted to 0.95 by the addition of humectants. Although the pH at the hot dog was adjusted with Glocono-delta-lacton to 5.4, the product had (p > 0.05) the least acceptance on account of the organoleptic changes (sour taste). Moreover, the temperature of 80 ± 1 °C for an hour with the aim of achieving an internal temperature of 75 °C was applied. Subsequently, the temperature of the hot dog samples reduced to around 5–6 °C within 40–45 min, and afterwards the sausages were kept at chilled temperature (>3 °C but 10 °C) throughout their shelf life. There was a decrease in total aerobic counts in hurdle treated hot dogs (with 50 ppm nitrite), compared to the control (with 120 ppm nitrite), whereas Clostridium perfringens counts and Clostridium botulinum detection were the same (p < 0.05) in both hurdle treated and control samples. The obtained results of present study clearly showed that both hurdle treated sample and control had the same (p < 0.05) overall acceptability and sensory attributes.     

Potential application of low dose gamma irradiation to improve the microbiological safety of fresh coriander leaves

A local market survey of fresh, raw coriander leaves, which is a component of many ready-to-eat, Indian dishes showed large variation in their bacterial load (7.0×10⁶–1.36×10⁸ cfu/gm) and mold and fungi (3.0×10³–1.0×10⁴ cfu/gm) and presence of potentially pathogenic bacteria such as faecal coliforms, Listeria and Yersinia. Conventional chlorination treatment affected the structural integrity of leaves leading to enhancement of rotting. Exposure to a low dose of 1 kGy was efficient for bacterial decontamination and elimination of potential pathogens without affecting the keeping quality of coriander leaves upto two weeks storage at 8–10 °C, measured visually in terms of degree of yellowing and rotting. Irradiation at higher doses led to the increased rotting during storage. The total chlorophyll contents as well its components, chlorophyll a and b did not change significantly on irradiation and subsequent storage. Similarly, the total carotenoid levels remained unaffected by exposure to 1 kGy dose, however, a dose dependent enhancement in their extractability was observed in irradiated leaves. No qualitative differences were observed in the GLC profile of volatile oils of control and irradiated (1 kGy) samples. There were no significant quantitative changes in the constituents of the major aroma compounds obtained from these two samples. Effects of irradiation on storage of the seasoning were also assessed.     

Determination of biogenic amines in sucuk

The biogenic amines content of sucuk (Turkish dry fermented sausage) were determined on 30 samples obtained from retail markets and butchers in Turkey. Also, pH and a_w values, microbiological properties, and NPN levels of these samples were analyzed. The most important biogenic amines were tyramine (range 2.4–676 mg/kg), followed by putrescine varied from not detected to 364 mg/kg. Histamine content was under 50 mg/kg in 80% of the samples while it was over 100 mg/kg in only one sample. Tryptamine was detected in 16 of 30 samples in the range of 1.2–82.3 mg/kg.

Lactic acid bacteria, Micrococcus/Staphylococcus and Enterobacteriaceae counts of samples varied from 2.84 to 8.85 log cfu/g, <2.00 to 6.74 log cfu/g, and <2.00 to 5.04 log cfu/g, respectively. pH values were varied between 4.53 and 6.29 while water activity (a_w) were measured to be between 0.761 and 0.960. NPN values were determined to be between 1.47% and 4.10%. Generally, total amine content was high in samples that had high NPN levels.     

SHALE OIL UPGRADING IN SUPERCRITICAL WATER SOLUTIONS

A supercritical water/HCl system was studied for upgrading Paraho shale oil (SO) at 400-425^°C. The key process parameters were water density (ρw) and HC1 loading. Increases in ρw increased the selectivity to distillate-range products at the expense of gas and coke products. Increases in ρw also enhanced the ability of HCl to catalyze N-removal. However, coke and gas make also increased with increases in HCl loading. Optimal or “best case” results and conditions were 897% N-removal and 75% distillate yield, at T = 425^°C, ρw = 0.2 g/cc, and HCl/SO = 0.05g/g. The underlying chemical mechanisms appeared to involve SCF solvent cage effects and hcterolytic N-removal brought about by dissociation of HCl.     

Microbiological safety of kinema, a fermented soya bean food

Kinema is a fermented soya bean food of Nepal and the hilly regions of North-eastern States of India. Generally, the fermentation is dominated by Bacillus spp. that often cause alkalinity and desirable stickiness in the product. The present study was undertaken in a limited number of commercial (market) kinema samples to test for the presence of foodborne pathogens and their properties. Bacillus cereus was present in numbers exceeding 10⁴ cfu/g product in five of the tested 15 market samples. Enterobacteriaceae and coliform bacteria exceeded 10⁵ cfu/g in 10 of the 15 samples. Escherichia coli exceeding 10⁵ cfu/g was found in two samples. Staphylococcus aureus was not detected in any of the tested samples. Of 31 isolated typical and atypical strains of B. cereus, 18 representative strains were tested qualitatively for the ability to produce diarrhoeal type enterotoxin (BCET) using an Oxoid BCET-RPLA test kit. Overall, BCET was formed by 12 strains in BHIG (brain heart infusion broth +1% glucose), by seven strains on sterilized cooked rice, and by five strains on sterilized cooked soya beans. Semi-quantitative tests on BCET revealed that levels exceeding 256 ng/g soya beans, produced by single pure culture inoculation with the isolated B. cereus strains, were reduced to ≤ 8 ng/g by frying kinema in oil, a common procedure when making kinema curry. It was also shown in a mixed pure culture experiment that a kinema strain B. Subtilis DK-W1, is able to suppress growth and BCET formation by a selected toxin producing strain (BC7-5) of B. cereus. It is concluded that the traditional way of making kinema and its culinary use in curries is safe. However, for novel applications of kinema, safety precautions are advisable.     

Microbiological quality of tomatoes and peppers produced under the good agricultural practices protocol AGRO 2-1 & 2-2 in Crete, Greece

The efficiency of the good agricultural practices (GAP) protocol AGRO 2-1 & 2-2, in advancing microbiological-quality of tomatoes and peppers, was studied in greenhouses at Ierapetra, Crete, Greece. The 240 tested vegetables-samples, produced under AGRO 2-1 & 2-2, showed satisfactory quality: Listeria monocytogenes absent per 25 g; Escherichia coli < 20 Colony Forming Units per gram (CFU/g); total coliforms 4.37–4.68 log CFU/g; aerobic plate counts 5.78–5.92 log CFU/g. Based on actual results and practices evaluation, we conclude that AGRO 2-1 & 2-2 can reduce microbial hazards for consumers and furthermore can establish practices in compliance to basic Euro-Retailer-Produce GAP (EUREPGAP) requirements.     

The in vitro antimicrobial and antioxidant activities of the essential oils and methanol extracts of endemic Thymus spathulifolius

The present study was conducted to evaluate the in vitro antimicrobial and antioxidant properties of essential oil and methanol extracts from a unique and endemic plant, Thymus spathulifolius (Hausskn. and Velen.). The antimicrobial test results showed that the essential oil of T. spathulifolius strongly inhibited the growth of test microorganisms studied, except for 4 fungi species while polar and non-polar subfractions of the methanol extract had moderate antibacterial, but not antifungal and anticandidal activity. The antioxidative potential of the samples was evaluated using two separate methods, inhibition of free radical 2,2-diphenyl-1-picrylhydrazyl (DPPH) and β-carotene–linoleic acid systems. The polar subfraction of the methanol extract was able to reduce the stable free radical 2,2-diphenyl-1-picrylhydrazyl (DPPH) with an IC₅₀ of 16.15 ± 0.5 μg/ml, which was lower than that of synthetic antioxidant, BHT, (19.8 ± 0.5 μg/ml). Inhibition values of linoleic acid oxidation were calculated as 92% and 89% for the oil and the polar subfraction, respectively. Gallic acid equivalent total phenolic constituent of the polar subfraction was 141.00 ± 0.90 μg/mg (14.1%, w/w). The chemical composition of a hydrodistilled essential oil of T. spathulifolius was analyzed by a GC and GC/MS system. A total of 28 constituents representing 99.2% of the oil were identified; thymol (36.5%), carvacrol (29.8%), p-cymene (10.0%) and γ-terpinene (6.3%) were the main components comprising 82.6% of the oil. Results presented here may suggest that the essential oil and extracts of T. spathulifolius possess antimicrobial and antioxidant properties, and therefore, they can be used as a natural preservative ingredient in food and/or pharmaceutical industry.     

Detection of Salmonella in food,equivalent to ISO 6579, by a three-days polymerase chain reaction-based method

A method is presented for the detection of Salmonella in food, which produces definitive results on the third day after the sample collection. The method is equivalent to ISO 6579 in terms of the same detection limit of 10⁰ cfu/25 g and 100% relative accuracy. The method involves pre-enrichment in buffered peptone water (16–20 h), two parallel selective enrichments in Rappaport–Vassiliadis and selenite–cystine medium, respectively (7 h), post-enrichment in Luria–Bertani medium (14–16 h), cell lysis by boiling in 1% Triton X-100, PCR using primers ST11, ST15 and a mimic internal control, and agarose gel electrophoresis. At the evaluation of the method on model food samples artificially contaminated with decimal dilutions of a Salmonella serotype Panama culture (cream cakes, ice cream, mayonnaise, meat products; 12 samples altogether), a detection limit of 10⁰ cfu/25 g was determined. This detection limit was confirmed by the evaluation of the method on the mentioned model food samples artificially contaminated with reference materials containing on average 5 cfu of S. Panama. At the evaluation of the method on naturally contaminated food samples (cream cakes, ice cream, mayonnaise, egg melange, minced and separated meat, ham and salami; 44 samples altogether), identical results (9 positives) as with the reference method were obtained.     

Elimination of Yersinia enterocolitica on carrots (Daucus carota L.) by using household sanitisers

In this study, the effects of lemon juice, the mixture of lemon juice–vinegar (1:1) and lemon dressing against Yersinia enterocolitica on carrot were investigated. Carrots inoculated with Y. enterocolitica cells (approximately 10⁶ cfu/ml) were treated with 100%, 75% and 50% lemon juice, a mixture of lemon juice–vinegar, and lemon dressing for 0, 15, 30 and 60 min. Enumeration of Y. enterocolitica populations on CIN agar beside TSA agar by using two methods as then performed. One hundred percent lemon juice was the most effective agent used, completely inhibiting viable cells of Y. enterocolitica on carrot after 15 min exposure. Although no growth was observed on carrots treated with 100% lemon juice for 15 min, there was no statistical difference between the antibacterial effects of 100% lemon juice, 75% lemon juice, the mixture of lemon juice–vinegar and lemon dressing whilst the effect of 50% lemon juice was statistically different from those solutions (P < 0.05) and its definite inhibitive activity was seen after 60 min treatment.     

Effect of usage Urtica dioica L. on microbiological properties of sucuk, a Turkish dry-fermented sausage

The effect of dried-ground Urtica dioica L. on microbiological characteristics of Turkish dry-fermented sausage (sucuk) were investigated. Dried-ground Urtica dioica L. was added to sucuk batters by the rate of 0%, 1%, 3% and 5%, and microbiological analysis were made on the 0, 3, 7, 10 and 14 days of ripening period.

According to the results, both the level of Urtica dioica L. and ripening time had a significant effects on the total aerobic mesophylic bacteria, lactic acid bacteria, Micrococcus/Staphylococcus, yeast-mould and Enterobacteriaceae counts (p<0.01). It was determined that the interactions ripening time × the levels of Urtica dioica L. had a significant effects on the counts of total aerobic mesophylic bacteria, lactic acid bacteria, Micrococcus/Staphylococcus, yeast-mould and Enterobacteriaceae (p<0.01). Enterobacteriaceae count were also found to be under the detectable level (<100 cfu/g) in sucuks (in day 14 of ripening) containing 3% and 5% Urtica dioica L.     

Occurrence of Vibrio spp. in sea fish, shrimps and bivalve molluscs harvested from Adriatic sea

One hundred and seventeen samples of sea fish (100), shrimps (10) and bivalve molluscs (7) were sampled at fish markets (100 samples) and hotels (17 samples) along the sea side in Croatia within the period from May to July 2000. The samples were tested for the presence of Vibrio spp. Vibrio spp. was isolated in 23 (19.65%) of total count of samples. The most frequently found were V. parahaemolyticus (9.40%), V. vulnificus (6.84%) and V. alginolyticus (3.42%). A significantly larger number of positive findings involved the samples collected at hotels (35.29%) as compared to those collected at fish markets (17.00%). The isolation was conducted by a conventional cultivation with the enrichment in alkaline peptone water and on TCBS Kobayashi agar (“Biolife”).     

Haematoporphyrin- and sodium chlorophyllin-induced phototoxicity towards bacteria and yeasts – a new approach for safe foods

Alternative methods for the decontamination of foods and surfaces in the food processing industry are under research. We introduce a new approach: the use of visible light plus photosensitizers, the photodynamic effect. First, we established an experimental model to examine the photosensitivity of immobilized micro-organisms. We tested subsequently several Gram-positive bacteria and yeasts for their photosensitivity after incubation with haematoporphyrin (HP) and sodium chlorophyllin (CHL). The survival levels of the bacteria fixed on solid media ranged from 0.0006% to 0.08% (3–5 log10 inactivations), and those of the yeasts from <0.1% to 50% (0.3–3 log10 inactivations) under the experimental conditions chosen.     

Microbiological characteristics of cake prepared from rice flour and sticky rice flour

Microbiological, chemical and sensory changes in MiGao (a traditional Chinese steamed cake) were studied during five days of storage at 25 °C. Microorganisms examined for were: Enterobactericeae, lactic acid bacteria, Gram-positive, catalase-positive cocci, Staphylococcus aureus, Bacillus strains, yeasts and moulds, and pH and moisture content were also analyzed. Total plate counts in the products under study in the third day of storage were in the range <10³ CFU/g and the shelf-life of the product was estimated to be only two or three days by the panelists. During the first two days, Gram-positive bacteria were dominant, mainly represented by Staphylococcus epidermidis. Bacillus strains occurred by the third day, reaching a maximum level of 1 × 10⁶ CFU/g after five days of storage. No Enterobactericeae or Lactic acid bacteria were detected in the processed products throughout the storage period. All the isolated strains from the mannitol salt agar belonged to the genus Staphylococcus. The predominant species were S. epidermidis and S. aureus. Most Bacillus strains, isolated from the tryptone soya agar, belonged to the species Bacillus brevis. The count of yeasts and moulds increased slowly but remained low throughout the storage period.     

Microbiological safety and quality of commercial sufu – a Chinese fermented soybean food

In this study, the microbiological safety and quality of commercial sufu were investigated. Twenty-three samples of three different types of sufu were obtained, mainly in China and some in The Netherlands. Chemical parameters analysed included moisture, pH, free amino N, NaCl, ethanol, sucrose, glucose, and fructose. Concentrations of NaCl, ethanol, glucose and fructose varied from 6.2%, 0.5%, 0% and 0% to 14.8%, 6.3%, 6.2% and 4.8%, respectively. Microbiological analyses were done for total count of mesophilic aerobic bacteria (TMAB), bacterial endospores, total count of halotolerant bacteria at 10% (THB10) and at 17.5% NaCl (THB17.5), lactic acid bacteria (LAB), fungi, Enterobacteriaceae, and the following pathogens: Bacillus cereus, Clostridium perfringens, Staphylococcus aureus and Listeria monocytogenes. High levels (>10⁵ CFU/g) of TMAB and bacterial endospores were found in most samples, and 85% of TMAB was identified as Gram-positive. Considerable levels (10⁵ and 10⁷ CFU/g) of LAB were detected in two samples of white sufu, and isolates of LAB were identified as most probably Lb. casei. One-third of the samples contained less then 10³ CFU/g B. cereus, but three samples had over 10⁵ CFU/g indicating potential hazard to consumers. All samples had less than 10³ CFU/g C. perfringens, except sample R11 (10⁵ CFU/g). S. aureus could not be detected in any of the samples tested since the competitive microflora (usually bacilli) disturbed typical features on the selective medium used; however S. aureus enterotoxin A was detected in some of the white and grey sufu samples. Fungi, Enterobacteriaceae, and L. monocytogenes were not detected in any of the samples. Based on these results, a microbiological guideline for safe commercial sufu is proposed.     

Survival of Listeria monocytogenes inoculated in retail soymilk products

The consumption of soymilk products has been growing; because these foods contain proteins and isoflavone, and lack of lactose and cholesterol. The survival and growth of Listeria monocytogenes in the soymilk products were investigated by inoculating Listeria monocytogenes into various retail soymilk products which claimed to improve or fortify the functional properties. The results show that the refrigeration cannot successfully prevent the growth of Listeria monocytogenes in soymilk products. The addition of fiber content and higher viscosity in soymilk may significantly inhibit the growth of L. monocytogenes at 4–7 °C; however, this phenomenon was not observed when the incubation temperature was elevated to 22 °C. The change of pH value and TTS (total soluble solids) in soymilk products owing to L. monocytogenes, especially those with rich fiber content, might be negligible for the public. Thus the possible contamination of L. monocytogenes to soymilk products should be of crucial concern.