In vitro effect of some fungicides on growth and aflatoxins production by Aspergillus flavus isolated from Capsicum powder

The aim of this study was to determine the effect of some pre-harvest fungicides on growth and aflatoxin (AF) production of three Aspergillus flavus strains found in Capsicum powder. Each isolate, previously isolated from paprika, chilli and smoked paprika, was inoculated on yeast extract sucrose agar and on a 3% paprika extract agar medium supplemented with different fungicides and incubated at 20 and 30°C during 7 days. Growth measurements were obtained on days 3, 5 and 7, and the AF production was determined on day 7. The significance of the effects of the factors (strain, medium, temperature, time and fungicides) and their interaction over colony diameter and AF production was determined. Temperature constrained the effectiveness of fungicides in reducing growth, the fungicides being most effective at 20°C. The efficacy of the fungicides over AF production depended on the medium used and temperature. The most effective fungicides in inhibiting growth and AF production, regardless of the strain tested or applied conditions, were tebuconazole 25% and mancozeb 80% applied at a concentration of 0.75 and 3.5 g l(-1), respectively. Care should thus be taken in the choice of a suitable fungicide because their effectiveness may depend on intra-specific variation and temperature. Moreover, it is necessary to take into account that the most efficient fungicide in reducing growth is not always the best choice for pre-harvest treatments because it may promote AF production. Thus, the best fungicide is the one that can simultaneous prevent growth and AF production.     

In vitro effect of some fungicides on growth and aflatoxins production by Aspergillus flavus isolated from Capsicum powder

The aim of this study was to determine the effect of some pre-harvest fungicides on growth and aflatoxin (AF) production of three Aspergillus flavus strains found in Capsicum powder. Each isolate, previously isolated from paprika, chilli and smoked paprika, was inoculated on yeast extract sucrose agar and on a 3% paprika extract agar medium supplemented with different fungicides and incubated at 20 and 30°C during 7 days. Growth measurements were obtained on days 3, 5 and 7, and the AF production was determined on day 7. The significance of the effects of the factors (strain, medium, temperature, time and fungicides) and their interaction over colony diameter and AF production was determined. Temperature constrained the effectiveness of fungicides in reducing growth, the fungicides being most effective at 20°C. The efficacy of the fungicides over AF production depended on the medium used and temperature. The most effective fungicides in inhibiting growth and AF production, regardless of the strain tested or applied conditions, were tebuconazole 25% and mancozeb 80% applied at a concentration of 0.75 and 3.5?g?l^?1, respectively. Care should thus be taken in the choice of a suitable fungicide because their effectiveness may depend on intra-specific variation and temperature. Moreover, it is necessary to take into account that the most efficient fungicide in reducing growth is not always the best choice for pre-harvest treatments because it may promote AF production. Thus, the best fungicide is the one that can simultaneous prevent growth and AF production.     

Microbiological studies on cassava fermentation for ‘lafun’ production

The microorganisms involved in the traditional fermentation of cassava for ‘lafun’ production were investigated. The microorganisms isolated during the fermentation include Bacillus sp., Leuconostoc sp., Klebsiella sp., Corynebacterium sp., Lactobacillus sp., Candida sp., Aspergillus sp., and Geotrichum sp. The moulds disappeared within the first 36h of fermentation. Bacillus sp. which were present at the beginning of fermentation decreased drastically as fermentation progressed. The yeasts appeared between 24–48h of the fermentation and increased rapidly. The lactic acid bacteria were implicated throughout the duration of the fermentation. Bacillus sp., Corynebacterium sp., Candida sp. and the lactic acid bacteria were considered to play important roles. The titratable acidity of the steeping water and tubers increased rapidly reducing the pH to below 4·0. The temperature of the steeping water varied between 30–32°C and usually not above the ambient. The moisture content of the fermenting tubers increased from 65% to 70% within the first 36 h and was never below 60% during the steeping period. The post-fermentation sun-drying reduced the moisture content to 12–15%.     

Selection and molecular characterization of wine yeasts isolated from the ‘El Penedès’ area (Spain)

A study of the microbiota present during the wine fermentation of five grape varieties from the ‘El Penedès’ area (Spain) was carried out to select autochthonous yeast strains for industrial wine production. In this study we identified members of the genera Candida, Dekkera, Hanseniaspora, Kluyveromyces, Torulaspora, Zygosaccharomyces and Saccharomyces in wine fermentation microbiota. Strains of Saccharomyces cerevisiae, as responsible agents of the alcoholic fermentation, were considered for a selection protocol. In this work we applied different enological criteria for selection, but previously we have characterized and differentiated Saccharomyces isolates by molecular methods to reduce the number of strains to analyse. Three strains were selected to conduct fermentations according to their characteristics. Finally, using mitochondrial DNA restriction analysis we demonstrated that the autochthonous selected strains are important contributors to the wine fermentation.     

Antibacterial potential of Enterococcus faecium strains isolated from ewes’ milk and cheese

The study was conducted to evaluate the antibacterial activity of three bacteriocin-producing Enterococcus faecium strains (TW15, TW20 and TW22) isolated from ewes’ milk and cheese sampled in the Patagonian region of Argentina. The strains were tested against spoilage and pathogens microorganisms showing antimicrobial activity towards 4 strains of Listeria monocytogenes, one strain of Listeria innocua and 2 strains of Staphylococcus aureus. E. faecium TW15, E. faecium TW20 and E. faecium TW22 were sensitive to vancomycin. Furthermore, investigation of virulence factors revealed the absence of the genes encoding them. The bacteriocin-like substances (BLISs) produced by the 3 strains were thermostable, pH resistant and can be expressed even in the presence of NaCl (3.0 g/100 g). Moreover, they prove to have a bactericidal mode of action. Results from physicochemical and biochemical characteristics of BLIS produced by these E. faecium strains make them potential candidates to aid in preservation of foods.     

Increased esters and decreased higher alcohols production by engineered brewer’s yeast strains

Esters and higher alcohols produced by yeast during the fermentation of wort have the greatest impact on the smell and taste of beer. Alcohol acetyltransferase, which is mainly encoded by the ATF1 gene, is one of the most important enzymes for acetate ester synthesis. Cytosolic branched-chain amino acid aminotransferase, on the other hand, which is encoded by the BAT2 gene, plays an important role in the production of branched-chain alcohols. The objective of this study is to construct engineered brewer’s yeast strains that produce more acetate esters and less higher alcohols. Industrial brewer’s yeast strain S5 was used as the parental strain to construct ATF1 overexpression and BAT2 deletion mutants. The engineered strains S5-2 and S5-4, which feature partial BAT2 allelic genes replaced by the constructed ATF1 overexpression cassette, were obtained. The ester production of the engineered strains was observed to increase significantly compared with that of the parental cells. The concentrations of ethyl acetate produced by the engineered strains S5-2 and S5-4 increased to 78.88 and 117.40 mg L^?1, respectively, or about 7.7-fold and 11.5-fold higher than that produced by parental S5 cells. The isoamyl acetate produced by S5-2 and S5-4 also increased to 5.14 and 9.25 mg L^?1, respectively; by contrast, no isoamyl acetate was detected in the fermentation sample of the parental strain S5. Moreover, S5-2 and S5-4, respectively, produced about 65 and 51 % of higher alcohols produced by the parental strain. The increase in acetate ester content and decrease in higher alcohol concentration shown by the engineered brewer’s yeast strains at the end of fermentation process indicate that the new strains are useful in future developments in the wheat beer industry.     

Quality of ‘Abacha’ from Fresh Cassava Roots and Dried Chips

This study was undertaken to determine the possibility of producing acceptable ‘abacha’ from dried chips. The ‘abacha’ slices were processed from dried cassava chips and fresh roots of four cassava varieties, TMS 98/2101, NR/87184, TMS 97/4779 and NR87184. The proximate composition, chemical and sensory properties as affected by the processing method and variety were evaluated. The proximate composition of the ‘abacha’ (on dry basis) showed moisture content range of 9.53 – 10.48%, protein 1.07 – 1.66%, ash 2.06 – 2.56%, fibre 1.72 – 1.95, fat 0.39 – 0.58% and carbohydrate content of 83.59 – 85.05%. The chemical composition of the wet ‘abacha’ slices showed pH of 5.60 – 5.80, total titratable acidity (TTA) of 0.047 – 0.063, and hydrogen cyanide (HCN) value of 7.80 – 10.41 mg/100 g. Generally, the TTA of the ‘abacha’ samples was not significantly (p > 0.05) affected by the method of processing ‘abacha’ and the variety used. Drying the wet ‘abacha’ reduced the pH to 5.20 – 5.30, increased the TTA to 0.213 – 0.230% and further reduced the HCN content to 7.32 – 8.63 mg/100 g. The HCN of all the ‘abacha’ samples processed from dried chips were significantly (p < 0.05) lower than the HCN of ‘abacha’ processed from fresh cassava root in some of the varieties. The lower sensory scores for ‘abacha’ made from dried chips shows that they were inferior to the ones from fresh cassava, although they were also accepted by the consumers. It also compared favourably with the ‘abacha’ processed from fresh cassava root in chemical qualities. This shows that when the need arises ‘abacha’ slices can be processed from dried cassava chips.     

Revisiting Fisher’s ‘Lady Tasting Tea’ from a perspective of sensory discrimination testing

The Lady Tasting Tea is a famous real story in the history of development of statistics, related to R.A. Fisher, one of the greatest statisticians and founders of modern statistics. The main learning and insight offered by this paper from revisiting the story are that the methodology of conventional sensory difference tests can be and should be expanded to cover the ‘M + N’ method with larger M and N. Unlike the conventional discrimination tests, which use multiple sets of ‘M + N’ samples with small M and N based on a binomial model, the ‘M + N’ tests with larger M and N can reach a statistical significance in a single trial using only one set of ‘M + N’ samples based on a hypergeometric distribution in Fisher’s exact test. This paper explores the applications of the new methods particularly in assessing performance of trained sensory panels and panelists. The connection of the odds ratio or common odds ratio with Cohen’s standardized mean difference d is also discussed.     

Characterization of exopolysaccharide produced by Lactobacillus kefiranofaciens ZW3 isolated from Tibet kefir – Part II

ZW3 is a newly discovered exopolysaccharide (EPS) produced by Lactobacillus kefiranofaciens ZW3, isolated from Tibet kefir. Some of its properties have been characterized in our previous paper. Present research demonstrates some other important aspects of this EPS. The molecular weight obtained by gel permeation HPLC was 5.5 × 10⁴ Da. Solubility, water holding and oil binding capacity of ZW3 EPS were 14.2%, 496.0%, and 884.74% respectively. Scanning electron microscopy (SEM) of ZW3 EPS demonstrated a smooth surface with compact structures. A topographical examination of EPS by atomic force microscopy (AFM) revealed that ZW3 EPS is composed of almost uniform net of molecules. Rheological study indicated that common salt did not affect the viscous behavior of ZW3 EPS and acidic pH may enhance its viscosity. Exopolymer showed a melting point of 93.38 °C. A degradation temperature (Td) of 299.62 °C was observed from the TGA curve for the polysaccharide ZW3.     

Sex-dependent modulation of immune development in mice by secretory IgA–coated Lactobacillus reuteri isolated from breast milk

Lactobacilli, commonly present in human breast milk, appear to colonize the neonatal gut and provide protection to infants against various infections, thereby promoting immune development. This study examined the potential probiotic role of breast milk–derived Lactobacillus reuteri FN041 in immune development in mice. The FN041 were gavaged either to BALB/c dams (n = 6/group) during the lactation period or to their offspring (n = 6/sex per intervention) after weaning separately (cointervention). All interventions induced increased intestinal barriers in 5-wk-old offspring, especially in the females. Immunoglobulin A plasmocytes in ileal tissue and secretory IgA (sIgA) in ileal contents increased in all 5-wk-old offspring of cointervention. The activation of mRNA expression of 17 genes was sex-dependent, especially in 5-wk-old offspring. Broader genes were regulated in female mice. The effect of cointervention on the Shannon index of total microbiota is sex-related. The Shannon index of sIgA-coated microbiota increased in both sexes. The sIgA-coated microbiota showed intergroup differences according to β diversity, especially in female mice that showed an increase in Bifidobacterium of Actinobacteria. The sIgA-coated Bifidobacterium was positively correlated with mRNA expression of Tlr9. The sIgA-coated Lactobacillus in male offspring was negatively correlated with mRNA expression of Cldn2. In conclusion, L. reuteri FN041 promoted the production of intestinal sIgA and the expression of genes related to antimicrobial peptides in the offspring and enhanced the function of the mucosal barrier, depending on sex and treatment manner.     

Optimization of β-carotene production by a mutant of the lactose-positive yeast Rhodotorula acheniorum from whey ultrafiltrate

The present work investigated on carotenogenesis with high β-carotene content by a new isolated high-activity strain-producer Rhodotorula acheniorum mutant MRN in cheese whey ultrafiltrate. After a serial of UV, ethymethanesurfonate (EMS), and nitrosoguanidine (NTG) mutagenesis, a mutant named MRN of the red lactose-positive yeast strain R. acheniorum was obtained. Then, the effects of different growth medium factors on carotenoid production by this mutant at batch-scale level were identified and optimized by means of response surface methodology (RSM) in order to achieve high-level production of β-carotene. The optimum conditions required to achieve the highest level of β-carotene (262.12±1.01 mg/L) were determined as follows: whey ultrafiltrate (WU) lactose concentration 55 g/L, pH 5.85, ammonium sulfate concentration 3.5 g/L, temperature 23°C, and aeration rate 1.56 vvm. The medium optimization resulted in a 6.45-fold increase in volumetric production (262.12±1.01 mg/L) and a 4.62-fold increase in the cellular accumulation (10.69±0.19 mg/g) of β-carotene.     

Technological characterization of lactic acid bacteria isolated from Armada cheese (a Spanish goats’ milk cheese)

Thirty-one strains of lactic acid bacteria (LAB) isolated from Armada cheese, Sobado variety, (eight strains of Lactococcus lactis subsp. lactis, four strains of Lactococcus lactis subsp. cremoris, two strains of L. lactis subsp. lactis biovar. diacetylactis, two strains of Leuconostoc mesenteroides subsp. mesenteroides, two strains of Leuconostoc mesenteroides subsp. dextranicum, five strains of Lactobacillus plantarum, six strains of Lactobacillus casei subsp. casei and two strains of Lactobacillus brevis) were screened for their acidifying capacity and enzymatic activity, that included the rapid API-ZYM system, the proteolytic activity, the amino-, di-, and carboxypeptidase activity and the caseinolytic activity. The strains of L. lactis subsp. lactis exhibited the highest acidifying and proteolytic activity. Lipase and esterase activity was practically non-existent for lactococci and lactobacilli; a certain esterase activity was observed among leuconostoc. The highest aminopeptidase activity was demonstrated by the cell-free extract (CFE) of some strains of L. plantarum, L. casei subsp. casei and L. mesenteroides subsp. dextranicum. The CFEs of L. lactis subsp. cremoris and L. lactis subsp. lactis possessed carboxypeptidase and dipeptidase activities, at levels depending on the strain. Appreciable caseinolytic activity was detected for the CFE of L. plantarum and those some lactococci.     

Radical Scavenging,Total Antioxidant Capacity,and Antiproliferative Activity of Phenolic Extracts from Extra Virgin Olive Oil by Cultivar ‘Frantoio’

The beneficial health effects of extra virgin olive oil are due to both its high content of monounsaturated fatty acids and its high content of phenolic compounds, which have recently attracted research interest. In this context, the aim of this work was to examine the in vitro antioxidant and antiproliferative activities of the phenolic extract obtained from extra virgin olive oil from O. europea cultivar ‘Frantoio’ (samples 1–4), one of the main varieties cultivated in Italy. The total phenolic content was determined by Folin–Ciocalteu method and instead the phenolic profile was obtained by HPLC coupled to a diode array detector and mass spectrometry. Extra virgin olive oil extracts exhibited high antioxidant activity through different mechanisms of action and these activities are related to the phenolic content. Sample 3 demonstrated the strongest free radical scavenging activity with IC₅₀ value of 56.5 μg/mL. The reducing ability measured with FRAP assay revealed that samples ranged from 91.3 to 156 μM Fe(II)/g. The same interesting trend was observed with Trolox equivalent antioxidant capacity value. Moreover, the virgin olive oils showed a good oxidative stability ranging between 19 to 32 h. Antiproliferative activity evaluated by SRB assay revealed that phenolic extracts from the cultivar ‘Frantoio’ showed a strong antiproliferative activity against CORL-23 cell line with an IC₅₀ value of 14.5 and 55.9 μg/mL for samples 3 and 1, respectively, and these results are comparable to the positive control vinblastine. Overall, these results showed that extra virgin olive oils from the cultivar ‘Frantoio’, may represent an interesting source of phenolic compounds as functional components that could be consumed in diets and/or used for the elaboration of functional food and pharmaceutical industries.