Effects of some Chinese medicinal plant extracts on five different fungi

Chinese medicinal plant extracts were screened against some fungal strains, such as Aspergillus niger, Botrytis cinerea, Fusarium moniliforme, Glomerella cingulata, and Phyllosticta caricae. Plants were extracted with hot water, 80% methanol or acetone. Aliquots of the extracts at variable concentrations were then incubated with different fungal strains, and the minimum inhibitory concentrations (MICs) of each plant extract determined. In this study, the methanol extracts of Cinnamomum cassia had MIC values of 13.3 mg ml⁻¹, when tested against F. moniliforme and P. caricae. The acetone extracts of C. cassia had MIC values of 8.3 mg ml⁻¹ and 10 mg ml⁻¹ respectively, when tested against B. cinerea and G. cingulata. The hot water extracts of C. cassia inhibited significantly the growth of A. niger, B. cinerea, F. moniliforme, and P. caricae with MIC values at 10, 11.7, 5, and 6.7 mg ml⁻¹ respectively. The acetone extracts of Curcuma longa inhibited effectively P. caricae with the MIC value at 6.7 mg ml⁻¹. To determine the stability, various plant extracts were stored at 4 and 25 °C over a period of one month and their effects on fungal growth examined. Results show that the acetone extracts of Cu. longa and Coptidis rhizoma maintained their activity against fungal strains when stored at 4 °C, but not at 25 °C. The methanol extracts of C. cassia lost a great portion of inhibitory activities but not all, after stored at 4 °C and 25 °C for one month. The effect of various combinations of these extracts on antimicrobial activity has also been examined. The combinations of herb extracts showed higher inhibitory effect towards tested fungi than that of individual extract. Results from these findings suggest that these herbal extracts may be used as natural antifungal agents to inhibit growth of foodborne pathogen.     

The dissipation rates of cyprodinil, fludioxonil, procymidone and vinclozoline during storage of grape juice

The disappearance of the fungicides cyprodinil, fludioxonil, procymidone and vinclozoline, which are widely used to control grey mold in vineyards, from commercially sterilized white grape juice was studied. Fungicide losses from white grape juice stored at 40 °C were monitored for about 2 months in order to simulate in a fast way the shelf-life of the product at room temperature (about 1 year). The fungicides were determined by using a simple method based on a liquid–liquid extraction (LLE) and gas chromatographic separation, followed by mass spectrometric detection (GC–MSD). Fortified white grape juice subsamples were taken three times a week from the oven and the degradation of the fungicides monitored for 56 days. The half-lives at 40 °C for vinclozoline and procymidone were found to be 11 and 20 days, respectively. Those for the more recently introduced fungicides fludioxonil and cyprodinil were somewhat longer (33 and 44 days, respectively). Based on the first-order rate constants obtained, the fungicides dissipated in the following sequence: vinclozoline (0.062 d⁻¹) > procymidone (0.035 d⁻¹) > fludioxonil (0.021 d⁻¹) > cyprodinil (0.016 d⁻¹).     

Determination of heavy metals in milk by potentiometric stripping analysis using a home-made flow cell

A method is described for sensitive and reliable determination of trace concentrations of cadmium, lead and copper in real samples by stripping potentiometry with a home-made flow cell. The selection of the experimental conditions was made using experimental design methodology. The optimum conditions for the method include an acetic acid–acetate buffer mixture (pH 3.4) as supporting electrolyte, an electrolysis potential of −1.1 V and a flow rate of 3 ml min⁻¹. The detection limits were 6.51 × 10⁻¹⁰ mol l⁻¹ for cadmium, 4.60 × 10⁻¹⁰ mol l⁻¹ for lead and 8.59 × 10⁻¹⁰ mol l⁻¹ for copper, with an electrolysis time of 900 s. The relative standard deviations at this concentration level were 0.038, 0.086 and 0.095, respectively. In order to check the analytical accuracy, standard solutions have been used and recoveries close to 100% were obtained. The method was successfully applied to the determination of cadmium, lead and copper in powdered milk, using the method of standard additions.     

Simple methods for rapid determination of sulfite in food products

Two simple methods for sulfite determination in food products were proposed. The modified para-rosaniline (PRA)-formaldehyde method showed a much broader linear range (0.05–5.0 mg l⁻¹ as SO₂) than the commonly used procedure for the sulfur dioxide detection in the atmosphere (0.05–1.0 mg l⁻¹ as SO₂). By using a standard reference color card, this method only needed 5 min to complete a test. The 5,5′-dithiobis(2-nitrobenzoic acid) (DTNB) method was another good alternative for rapid determination of sulfite since it only needed one main reagent and the method was robust and easy to operate with the linear range of 0.10–4.3 mg l⁻¹ as SO₂. When applying to real food samples, the DTNB method had good recoveries for all the tested samples and the results agreed well with those obtained by the official iodometric titration. The modified PRA-formaldehyde method worked well with all the tested dried food products, though matrix effect of sulfite binding was observed for the beer samples. Thus, the modified PRA-formaldehyde method has advantages of sensitivity and rapidity, but the DTNB method offers a wider range of applications. Both methods provided practical ways for in situ determination of sulfite by non-professional operators with the modified PRA-formaldehyde method suited for food samples with less sulfite binding problems.     

Determination of cadmium and lead in fruit juices by stripping chronopotentiometry and comparison of two sample pretreatment procedures

A method for the determination of cadmium and lead in apple and pear fruit juices by stripping chronopotentiometry (SCP) is described. The results obtained after previous digestion of the sample with concentrated sulphuric acid and dry-ashing (sample pretreatment procedure “A”) and after a treatment with concentrated hydrochloric acid followed by heating in hot water bath (sample pretreatment procedure “B”) were compared. The sample pretreatment procedures were tested on real fruit juice samples and all data confirmed that the efficiency of procedure A was greater than procedure B. In particular, for samples treated with procedure A, good linearity was obtained in the range of examined concentration as is shown by the determination coefficients that were 0.998 (n = 4) for cadmium and 0.996 (n = 4) for lead. Recoveries of 86–104% for cadmium and of 87–102% for lead were obtained from a sample spiked at different levels. The accuracy was also evaluated by means of a matching reference sample of spiked skim milk powder (BCR 150—Community Bureau of Reference) to prove the reliability of the method. The detection limits were 2.0 ng g⁻¹ for cadmium and 4.8 ng g⁻¹ for lead. The relative standard deviations (mean of nine determinations), evaluated on a real sample, were 7.8% and 6.5% respectively. The average content was in the range not detectable–3.0 ng g⁻¹ for cadmium and 8.2–21.3 ng g⁻¹ for lead.     

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.     

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.     

Analysis of 140 pesticides from conventional farming foodstuff samples after extraction with the modified QuECheRS method

Pesticides in foodstuff are becoming a major issue due to their intensive use in agriculture. Thus an appropriate control of their residues in food samples has to be operated. In this study we analysed 105 pesticides with GC/SQ-MS and 46 pesticides with HPLC/IT-MS after extraction with the QuECheRS method in four matrices (grape, lemon, onion and tomatoes). For GC-amenable substances, the LOD and LOQ ranged from 0.4 to 48.2 μg/kg and from 1.2 to161 μg/kg, respectively. For HPLC-amenable substances, they varied from 1.0 to 115 μg/kg and from 3.3 to 382 μg/kg, respectively. With GC/MS, 61–82% of the substances showed a recovery in the range of 70–110% and 6–30 % presented a recovery higher than 110% at the 500 μg/kg fortification level. With HPLC/MS, 87–93% of the substances presented recoveries in the range of 70–110% at the 500 μg/kg fortification level compared to 78–85% at the 50 μg/kg fortification level. Lemon and onion showed poor recoveries but are known to be difficult matrices (high acidic and high sulfur content, respectively). The method was proved to be repeatable with RSD lower than 20% at 500 μg/kg and lower than 15% at 50 μg/kg with both devices.     

Inhibition of food poisoning and pathogenic bacteria by Lactobacillus plantarum strain 2.9 isolated from ben saalga, both in a culture medium and in food

The bacteriocinogenic strain Lactobacillus plantarum 2.9 isolated from the traditional pearl millet-based African fermented food ben saalga was tested for inhibition of food poisoning and pathogenic bacteria in MRS broth and in a malted millet flour slurry. In MRS broth, strain 2.9 completely eliminated Bacillus cereus, Escherichia coli O157:H7 and Salmonella enterica cells within 48 h incubation at 22–30 °C. A much lower inhibition was observed at 15 °C. The inhibitory effect of strain 2.9 on the above-mentioned target bacteria was corroborated in the malted millet flour slurry, reducing viable cell counts below detection levels after 8 h storage for B. cereus or after 24 h for S. enterica and 48 h for E. coli. An Enterobacter aerogenes strain was only moderately inhibited in the slurry. Results from the present study suggest that strain 2.9 could be used as starter culture to improve the microbiological safety of fermented ben saalga.     

Incidence of aflatoxins, ochratoxin A and zearalenone in tunisian foods

A total of 209 samples of different groups of foods widely consumed by the Tunisian population were collected during 2004–2005 years. Samples were analyzed for contamination with aflatoxins, ochratoxin A and zearalenone, using competitive enzyme-linked immunosorbent assay (ELISA). The predominant mycotoxin was ochratoxin A with a mean level of 3.5 ± 5.3 ng g⁻¹ in 59.8% of studied samples. Furthermore, Aflatoxins were detected in all analyzed commodities with a contamination frequency of 50.5%. In addition, aflatoxin B1 was found in 37% of the samples. The zearalenone was detected around 15% with a mean level of 10.4 ± 11.8 ng g⁻¹. Species, dried fruits and sorghum were the most contaminated samples by aflatoxin and ochratoxin mycotoxins, whereas Rice was the least contaminated commodity. The most frequent mycotoxins co-occurrence included aflatoxins and ochratoxin A, which have been detected in 33.8% of analyzed samples. Furthermore, the simultaneous contamination by aflatoxins, ochratoxin A and zearalenone was observed in 7.2% of studied samples.     

Occurrence of pathogenic bacteria in raw milk, cultured pasteurised milk and naturally soured milk produced at small-scale dairies in Zimbabwe

The presence of selected pathogenic bacteria in raw milk (RM), cultured pasteurised milk (CPM), and naturally soured raw milk (NSRM) sold at three small-scale dairies in Zimbabwe was determined. Mean levels of Escherichia coli in RM, CPM and NSRM were 4.5, 7.1 and 7.8 log₁₀ CFU ml⁻¹, respectively. Enterotoxigenic E. coli (ETEC), producing heat-stable enterotoxin (ST1) was found in 16 of the samples. Presumptive Staphylococcus aureus was identified in 7 of 12 RM, 15 of 27 CPM and 20 of 21 NSRM samples at mean levels of 5.2, 7.3 and 7.8 log₁₀ CFU ml⁻¹, respectively. Klebsiella pneumonia spp., Enterobacter cloacae, Proteus mirabilis, Hafnia alvei, Citrobacter spp., Serratia marcescens and Aeromonas hydrophilia were found in 27 samples. The high number of S. aureus, E. coli, identification of ETEC-ST1 and the other pathogenic microorganisms found in pasteurised and unpasteurised milk products represent a health hazard to the consumers and emphasises the need for improved hygiene practice at all levels in the dairy.     

Molecular grouping and pathogenic analysis of Listeria monocytogenes of clinical and food origin

The phylogenesis and pathogenesis of ten Chinese Listeria monocytogenes isolates of clinical (n = 2) and food (n = 8) origin have been investigated in this study. The 597 bp nucleotide sequence at 3′ terminal of actA gene of these L. monocytogenes isolates were amplified and sequenced. Compared with those of published sequences of the corresponding region within the same gene, the phylogenetic tree was constructed using MEGA2.1 software. It was shown that one clinical strain 1579 and food strains YZ5, YZ7, YZ8 belong to lineage I, while the other clinical strain 1191, and food strains YZ1, YZ2, YZ3, YZ4, YZ6 belong to lineage II. The virulence of these strains based on experimental infection model of SPF chick embryos were determined, and compared between L. monocytogenes isolates of two lineages and their hemolytic activity was assessed according to the development of zones of hemolysis around the colonies. Isolates of lineage I showed high virulence while isolates of lineage II (except clinical isolates 1191) exhibited low virulence, since 100% mortality rate and relative short time to death for embryos were observed for strains of lineage I, while 20–30% mortality rate and long time to death for embryos were observed for isolates of lineage II. Hyper-virulent isolate 1579, YZ8 and lower-virulent isolates YZ2, YZ3, YZ4 and YZ6 show strong hemolytic activity while hyper-virulent isolate 1191, YZ5, YZ7 and lower-virulent isolate YZ1 show weak hemolytic activity. The discrepancy between virulence and hemolytic activity indicates that L. monocytogenes hemolytic activity is not directly proportional to their virulence and thus it can not be used as a reliable criteria for assessment of the virulence of L. monocytogenes.     

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 Campylobacter jejuni on vacuum or carbon dioxide packaged primal beef cuts stored at −1.5 °C

The ability of two Campylobacter jejuni strains (MOO3, a cattle isolate, and MOO8, a sheep isolate) to survive on chilled, preservatively packaged primal beef cuts was examined. Each of the strains was inoculated separately (10⁵ cfu g⁻¹) onto 500 g beef steaks, packaged under vacuum or 100% carbon dioxide, and stored, with uninoculated controls, for 41 days at −1.5 °C. Bacterial numbers were determined at 0, 1, 3, 6, 13, 20, 27, 34 and 41 days by dilution, plating on both Columbia Sheep Blood Agar and Campylobacter Blood-Free Selective Agar and incubation at 42 °C for 48 h under microaerophilic conditions. In addition, an aerobic count on all samples was determined by dilution, plating on Plate Count Agar and incubation at 25 °C for 72 h. No significant changes in numbers of the C. jejuni strains or aerobic bacteria occurred during storage at −1.5 °C on steaks packaged under either vacuum or carbon dioxide. The ability of these pathogens to survive standard preservative packaging conditions is different from that reported in the literature and therefore a cause for concern. Strict hygienic practice or the implementation of decontamination technologies are recommended as mechanisms to assure safety of meat with respect to this pathogen.     

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.     

Antibacterial activities of the surface microflora of Kefalograviera cheese

The microflora developed on the surface of Kefalograviera, a hard cheese, during ripening in the warm room was studied. The microbial total counts and yeasts decreased (P < 0.05) and the same was observed with the pH on ripening for 15 days. Bacilli dominated at day 4 while at 15 days Micrococcaceae and enterococci occurred frequently. A great number (38 out of 56) of the isolates exhibited narrow spectrum of antibacterial activities against Gram-positive indicator strains, while ten of the Bacillus isolates (Bacillus pumilus) inhibited the growth of Listeria monocytogenes. The extracellular antilisterial substances were sensitive to proteinase K and heat labile (at 100 °C).     

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.     

An Improved Desulfurization Process Based on H2O2/Formic Acid Oxidation and Liquid-Liquid Extraction 2. FCC Diesel Oil Feedstocks

Semi-empirical molecular orbital theory and parametric model 3 methods were used to calculate the quantum chemistry properties of dibenzothiophene (DBT) and its corresponding sulfone compounds to investigate the essential theory of a formic acid/H₂O₂ oxidation desulfurization system. To study oxidation kinetics of the oxidation desulfurization reaction, DBT was also employed as a model compound to obtain the oxidation kinetic parameters. Desulfurization of fluid catalytic cracking (FCC) diesel was also performed in this process, and the sulfur content was reduced from 7,200 μg·g^-1 to 598 μg·g^-1 after being extracted by polyethylene glycol 200.     

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.     

Assessment of estimated daily intake of nitrite by average consumption of processed foods in Korea

This study has been performed to estimate the average daily intake of nitrite used in Korea as a color fixative. The crude estimation of daily intake was calculated based on maximum permitted levels (MPL) and national food disappearance data in 1998. In order to refine estimated daily intake (EDI), daily food consumption nationwide National Health and Nutrition Survey in 1998 and the concentration of nitrite in their permitted foods were applied. The crude EDI of nitrite was 17.85 μg/kg bw/day, representing 25.5% of acceptable daily intake (ADI) assigned by JECFA. The refined average EDI for nitrite was 0.87 μg/kg bw/day, representing 1.25% of ADI. For average consumers of age–sex groups ranged from 0.2 to 4.8 μg/kg bw/day, representing 0.3%–6.9% of the ADI.