A complex microbial consortium derived from raw milk and composed of populations classified in 4 groups (lactic acid bacteria (A), Gram positive catalase positive bacteria (B), Gram negative bacteria ...(C) and yeasts (D)) can contribute to the inhibition of
Listeria monocytogenes in the core of an uncooked pressed cheese. To identify what groups may be involved in the inhibition, the consortium was simplified by successively omitting one group at a time. Pasteurized milk was inoculated with these more or less complex consortia and their effects on
L. monocytogenes count, pH, acids and volatile compounds in the core of uncooked pressed cheese were evaluated. The growth of
L. monocytogenes was the highest in cheeses prepared with pasteurized milk and only
St. thermophilus. Inhibition in other cheeses was expressed by comparison with growth in these ones. All the consortia containing both lactic acid bacteria (group A) and Gram positive catalase positive bacteria (group B) — ABCD, ABD, ABC, AB — were more inhibitory than those containing lactic acid bacteria on its own (A) or associated only with yeasts (AD) or/and Gram negative (ADC). Consortia without lactic acid bacteria were weakly inhibitory or had no effect. Gram positive catalase positive bacteria alone were not inhibitory although most of the species became established in the cheeses. The
Lactobacillus population (
Lb. casei,
Lb. plantarum,
Lb. curvatus and
Lb. farciminis) was predominant in cheeses (9
log
CFU/g) with a higher count than
Leuconostoc (7
log
CFU/g) and
Enterococcus (7
log
CFU/g).
Lactobacillus counts were negatively correlated with those of
L. monocytogenes (r
=
−0.84 at 18
days) and with the level of
d-lactic acid. There was no correlation between
L. monocytogenes and
Leuconostoc or
Enterococcus counts. Complex consortium ABCD and AB not only had a stronger inhibitory power in cheeses than consortium AD, they were also associated with the highest levels of
l-lactic and acetic acids. All cheeses inoculated with lactic acid bacteria differed from those without by higher levels of ethyl formiate, pentane and alcohols (2-butanol, 2-pentanol), and lower levels of ketones (2-hexanone, 2,3-butanedione) and aldehydes (2-methyl-butanal). Levels of 2-methyl-butanal, 2-butanol and 2-pentanol were higher in ABCD and AB cheeses than in AD cheeses. Beside their contribution to the inhibition, their effect on cheese flavour must be evaluated.
► Methodology for simplifying a complex antilisterial consortium in cheese.► Synergy of lactic acid and ripening bacteria in the inhibition. ► Dominance of
Lactobacillus producing lactic acid. ► Higher levels of acetic acid and alcohols in inhibitory cheeses.
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GEOZS, IJS, IMTLJ, KILJ, KISLJ, NUK, OILJ, PNG, SAZU, SBCE, SBJE, UL, UM, UPCLJ, UPUK
The microbial basis of acidification process during spontaneous cheese whey wastewater fermentation was decrypted by implementing both culture-dependent and culture-independent techniques. Lac ...tobacillus and Bifidobacterium were the predominant taxa among the microbiota growing on MRS (deMan, Rogosa, and Sharpe), while Kazachstania unispora and Dekkera anomala yeast species were also isolated. Almost all Lactobacillus isolates were heterofermentative that could ferment glucose and lactose, with most of them being related to Lactobacillus hilgardii (99.0–100 % similarity). By employing fluorescence techniques, the dominance of long crescent-shaped bacteria in the acidogenic sludge was observed. Temperature gradient gel electrophoresis (TGGE), clone library, and next-generation sequencing techniques revealed the dominance of Selenomonas lacticifex. Based on Illumina data, Selenomonas in the continuous stirred-tank reactor (CSTR) represented 70.13 ± 4.64 % of the bacterial reads, while other Veillonellaceae taxa (Megasphaera and Pectinatus) represented a notable proportion (6.54 %). Prevotella was only detected by Illumina sequencing as an important constituent of the microbial population (14.97 ± 1.71 %). Budding yeasts represented 97 % of the fungal population in the CSTR, with Yarrowia strains representing 88.85 ± 5.52 % of the fungal reads. Spontaneous cheese whey acidification can favor the dominance of rumen bacteria and here was driven by the rarely reported S. lacticifex-type fermentation, which should be taken into consideration during evaluation of acidogenesis in process simulation and modelling. Moreover, the important nervonic acid content detected indicates that acidogenic sludge can be used as a source for the production of high value-added biomedical substrates.
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CEKLJ, DOBA, EMUNI, GEOZS, IJS, IMTLJ, IZUM, KILJ, KISLJ, MFDPS, NUK, OILJ, PILJ, PNG, SAZU, SBCE, SBJE, SBMB, SBNM, UILJ, UKNU, UL, UM, UPUK, VKSCE, ZAGLJ
The aim of the present study was to develop a predictive model to quantify the spoilage of yoghurt with fruits. Product samples were stored at various temperatures (5–20 °C). Samples were subjected ...to microbiological (total viable counts, lactic acid bacteria-LAB, yeasts and moulds) and physico-chemical analysis (pH, titratable acidity and sugars). LAB was the dominant micro-flora. Yeasts population increased at all temperatures but a delay was observed during the first days of storage. Titratable acidity and pH remained almost constant at low temperatures (5 and 10 °C). However, at higher temperatures (>10 °C), an increase in titratable acidity and reduction in pH was observed. Sugar concentration (fructose, lactose and glucose) decreased during storage. A mathematical model was developed for shelf-life determination of the product. It was successfully validated at a temperature (17 °C) not used during model development. The results showed that shelf-life of this product could not be established based only on microbiological data and use of other parameters such as sensory or/and physico-chemical analysis is required. Shelf-life determination by spoilage tests is time-consuming and the need for new rapid techniques has been raised. The developed model could help dairy industries to establish shelf-life predictions on yoghurt with fruits stored under constant temperature conditions.
► Practical use by food industry of kinetic growth models which determine food shelf-life. ► Predictive model able of predicting yogurt shelf-life under constant temperature conditions. ► Information on shelf-life of specific dairy products category. ► Useful and practical tool for quality monitoring by dairy industry. ► Assistant in predicting storage life of yogurts with fruits under concrete business conditions.
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GEOZS, IJS, IMTLJ, KILJ, KISLJ, NUK, OILJ, PNG, SAZU, SBCE, SBJE, UL, UM, UPCLJ, UPUK
•We studied the effect of packaging with citrus extract and natamycin on a deli salad.•Addition of citrus extract improved the taste and odour of salad kept under vacuum.•Salad’s shelf-life in air ...was extended (>10days) using citrus extract–natamycin.•The presence of citrus extract did not negatively influence the taste of salad.
The effect of packaging (aerobic and vacuum) either with citrus extract, natamycin individually added, or the combination of two, was studied on the shelf-life of a Greek traditional deli salad “Tzatziki” during storage under refrigeration (4°C). Irrespective of the packaging conditions and treatments, yeasts populations of approximately 4.0–6.5logcfu/g were recorded in the microflora of the salad, whereas the Pseudomonas spp. populations were lower (2–3logcfu/g). Tzatziki’s overall flavour was better under vacuum, and of all the treatments examined, the addition of citrus extract, and to a letter extent the combination with natamycin, improved the taste and odour (fruity, pleasant, refreshing with reduced garlic typical flavour) of Tzatziki salad. The shelf-life of Tzatziki was extended by ca. >10days (citrus extract, citrus–natamycin) and 5–6days (natamycin, citrus and citrus–natamycin) under aerobic or vacuum, respectively, as compared to the control sample.
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GEOZS, IJS, IMTLJ, KILJ, KISLJ, NUK, OILJ, PNG, SAZU, SBCE, SBJE, UL, UM, UPCLJ, UPUK
Most transcription factors fulfill their role in protein complexes. As a consequence, information about their interaction capacity sheds light on a protein's function and the molecular mechanism ...underlying this activity. The yeast two-hybrid GAL4 (Y2H) assay is a powerful method to unravel and identify the composition of protein complexes. This in vivo based system makes use of two functional protein domains of the GAL4 transcription factor, each fused to a protein of interest. Upon interaction between the two proteins under study, a transcriptional activator gets reconstituted and reporter genes get activated, allowing the yeast to grow on selective medium. In this chapter protocols are given for Y2H library screening, directed Y2H screening, Y2H matrix screening, and YnH screening involving more than two proteins.
The performance of 3 blood culture bottles (BACTEC Plus Aerobic/F, Plus Anaerobic/F, and Anaerobic Lytic/F) were analyzed with clinical specimens collected from 688 Emergency Department patients. A ...total of 270 strains belonging to 33 species were identified, with E. coli and S. aureus as the most frequently detected. Overall recovery rate (RR) of bacteria and yeast was equivalent in the Plus Aerobic/F vials (208 of 270 isolates; 77.0%) and Anaerobic Lytic/F vials (206 isolates; 76.3%) and significantly better than in the Plus Anaerobic/F vials (189 isolates; 70.0%). Median time to detection (TTD) was earliest with the Anaerobic Lytic/F vials (12.0h) compared with the Plus Aerobic/F (14.6h) and Plus Anaerobic/F vials (15.4h). Positivity rate (PR) was similar for Anaerobic Lytic/F vials (76.9%) and Plus Aerobic/F vials (76.5%), but better if compared with Plus Anaerobic/F vials (69.4%). The PR and TTD for the combination of Plus Aerobic/F with Anaerobic Lytic/F (94.5% and 12.3h, respectively) was significantly better than with Plus Aerobic/F with Plus Anaerobic/F (87.8% and 14.1h).
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GEOZS, IJS, IMTLJ, KILJ, KISLJ, NUK, OILJ, PNG, SAZU, SBCE, SBJE, UL, UM, UPCLJ, UPUK
This article reports on the investigation on the bioactivity of eugenol, limonene, and citrus extract against three bacteria (Lactobacillus plantarum, Lactobacillus brevis, and Bacillus coagulans) ...and three yeasts (Saccharomyces bayanus, Pichia membranifaciens, and Rhodotorula bacarum), representing the spoilage microflora of fruit juices. The experiments were performed with laboratory media by using a microdilution method. Data were fitted using the Gompertz equation, and the kinetic parameters were used to evaluate the MIC and the dose-dependent effect (at suboptimal doses for each essential oil). Citrus extract was the most effective essential oil, and the results suggested the following susceptibility hierarchy, from the most sensitive microorganism to the most resistant one (values in parentheses represent MICs): S. bayanus (2 ppm) > R. bacarum (3 ppm) > P. membranifaciens (5 ppm) > B. coagulans (cells, 20 ppm) > L. brevis (40 ppm) > L. plantarum (>40 ppm).
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CEKLJ, GEOZS, IJS, IMTLJ, KILJ, KISLJ, NLZOH, NUK, OILJ, PNG, SAZU, SBCE, SBJE, UILJ, UL, UM, UPCLJ, UPUK, ZAGLJ
The non-conventional yeast Arxula adeninivorans uses 1-butanol as a carbon source and has recently attracted attention as a promising organism for 1-butanol production. Alcohol dehydrogenases (adhp) ...are important catalysts in 1-butanol metabolism, but only Aadh1p from Arxula has been characterized. This enzyme is involved in ethanol synthesis but has a low impact on 1-butanol degradation.
In this study, we identified and characterized a second adhp from A. adeninivorans (Aadh2p). Compared to Saccharomyces cerevisiae ADHs' (ScAdh) protein sequences it originates from the same ancestral node as ScAdh6p, 7p and 4p. It is also localized in the cytoplasm and uses NAD(H) as cofactor. The enzyme has its highest activity with medium chain-length alcohols and maximum activity with 1-butanol with the catalytic efficiency of the purified enzyme being 42 and 43,000 times higher than with ethanol and acetaldehyde, respectively. Arxula adeninivorans strain G1212/YRC102-AADH2, which expresses the AADH2 gene under the control of the strong constitutive TEF1 promoter was constructed. It achieved an ADH activity of up to 8000 U/L and 500 U/g dry cell weight (dcw) which is in contrast to the control strain G1212/YRC102 which had an ADH activity of up to 1400 U/L and 200 U/g dcw. Gene expression analysis showed that AADH2 derepression or induction using non-fermentable carbon-sources such as ethanol, pyruvate, glycerol or 1-butanol did occur. Compared to G1212/YRC102 AADH2 knock-out strain had a slower growth rate and lower 1-butanol consumption if 1-butanol was used as sole carbon source and AADH2-transformants did not grow at all in the same conditions. However, addition of the branched-chain amino acids leucine, isoleucine and valine allowed the transformants to use 1-butanol as carbon source. The addition of these amino acids to the control strain and Δaadh2 mutant cultures had the effect of accelerating 1-butanol consumption.
Our results confirm that Aadh2p plays a major role in A. adeninivorans 1-butanol metabolism. It is upregulated by up to 60-fold when the cells grow on 1-butanol, whereas only minor changes were found in the relative expression level for Aadh1p. Thus the constitutive overexpression of the AADH2 gene could be useful in the production of 1-butanol by A. adeninivorans, although it is likely that other ADHs will have to be knocked-out to prevent 1-butanol oxidation.
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IZUM, KILJ, NUK, PILJ, PNG, SAZU, UL, UM, UPUK
Diploid yeast, like most eukaryotes, can undergo meiotic differentiation to form haploid gametes. Meiotic differentiation and cell growth (proliferation) are mutually exclusive programs, and in yeast ...the switch between growth and meiosis is controlled by nutritional signals. The signaling pathways that mediate nutritional controls on meiotic initiation fall into three broad classes: those that respond to nutrient starvation, those that respond to non-fermentable carbon sources, and those that respond to glucose. At the onset of meiosis, nutritional signaling pathways converge on transcriptional regulation of two genes: IME1, which encodes a transcription factor; and IME2, which encodes a protein kinase. Transcription of IME1 and IME2 trigger initiation of meiosis, and the expression of these two genes is linked with one other, with expression of later meiotic genes and with early meiotic events such as DNA replication. In addition, the signaling pathways that control IME1 and IME2 expression are themselves integrated through a variety of mechanisms. Thus the signal network that controls the switch from growth to meiotic differentiation provides a signaling code that translates different combinations of extracellular signals into appropriate cellular responses.
Effect of fat replacers on kefir quality Ertekin, Bilge; Guzel-Seydim, Zeynep B
Journal of the science of food and agriculture,
15 March 2010, Volume:
90, Issue:
4
Journal Article
Peer reviewed
Open access
The purpose of the study was to determine the effects of fat replacers on the quality of non-fat kefir. Skim milk fortified with Dairy Lo® (DL) and inulin (INU) was fermented with kefir grains to ...manufacture kefir. The results of compositional, microbiological, rheological and sensorial analyses were compared with whole kefir (WK) and non-fat kefir (NFK) controls. Results for dry matter, pH and lactic acid ranged between 82.4 and 109.1 g kg⁻¹, 4.26 and 4.40, and 7.0 and 9.2 g L⁻¹, respectively. Acetaldehyde and ethanol contents of samples were between 2.89 and 7.28 mg L⁻¹, and 151.46 and 323.89 mg L⁻¹, respectively. In all samples, Lactobacillus spp., Streptococcus spp. and yeast counts were between 9.1 and 9.9, 9.3 and 9.9, and 5.2 and 5.6 log cfu mL⁻¹, respectively. Kefir samples had non-Newtonian behaviour and pseudoplastic fluid with thixotropy. At the first day, DL had the highest apparent viscosity (3.119 Pa s) while NFK had the lowest value (1.830 Pa s). In the sensory evaluation, odour and taste scores of samples were not different. Dairy Lo® and inulin could be used without any adverse effect for the production of non-fat kefir.
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BFBNIB, FZAB, GIS, IJS, KILJ, NLZOH, NUK, OILJ, SBCE, SBMB, UL, UM, UPUK
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