Evaluation of culture media for enumeration of Lactobacillus acidophilus, Lactobacillus casei and Bifidobacterium animalis in the presence of Lactobacillus delbrueckii subsp bulgaricus and Streptococcus thermophilus

The study compared the growth capability of probiotic (Lactobacillus acidophilus La05, Lactobacillus casei Lc01 and Bifidobacterium animalis Bb12) and non-probiotic (Lactobacillus delbrueckii subsp bulgaricus and Streptococcus thermophilus) cultures on twenty-one culture media grouped according to selectivity: non-selective agars, selective agars without antibiotics and MRS agars containing different combinations of lithium chloride, cystein, bile salts and antibiotics. Four of these media were selected for quantitative enumeration of L. acidophilus La05, L. casei Lc01, and B. animalis Bb12. The best culture media and incubation conditions for enumeration of the probiotic cultures were: B. animalis: MRS agar with dicloxacillin, 37 °C or 42 °C, anaerobiosis; L. acidophilus: MRS agar with bile salts, 37 °C or 42 °C, aerobiosis; L. casei: MRS agar with lithium chloride and sodium propionate, 37 °C or 42 °C, aerobiosis or anaerobiosis. Plating on MRS with glucose replaced by maltose, 37 °C or 42 °C, anaerobiosis, will distinguish probiotic from non-probiotic cultures. For enumeration of each probiotic in a mixed culture, the following media and incubation conditions were recommended: B. animalis: 4ABC-MRS, 42 °C, anaerobiosis, L. acidophilus: LC medium, 42 °C, aerobiosis or anaerobiosis and L. casei: LP-MRS, 42 °C, aerobiosis or anaerobiosis. In all experiments, differences in counts using pour plating or surface plating were not significant (P ≤ 0.05).     

Survival of Lactobacillus acidophilus and Bifidobacterium animalis ssp. lactis in stirred fruit yogurts

The effect of commercial fruit preparations (mango, mixed berry, passion fruit and strawberry) on the viability of probiotic bacteria, Lactobacillus acidophilus LAFTI^® L10 and Bifidobacterium animalis ssp. lactis LAFTI^® B94 in stirred yogurts during storage (35 days) at refrigerated temperature (4 °C) was evaluated. The results showed that addition of either 5 or 10 g/100 g fruit preparations had no significant (p>0.05) effect on the viability of the two probiotic strains except on L. acidophilus LAFTI L10 yogurt with 10 g/100 g passion fruit or mixed berry. After the addition of fruit preparation, 96% of the yogurts incorporated with fruit preparation did not exhibit a greater loss in the viability of probiotic bacteria compared to plain yogurt during the storage period. A correlation between the post-storage pH in yogurts and the survival of probiotic bacteria was observed. All the yogurts, however, contained the recommended levels of (10⁶-10⁷ cfu/g) probiotic bacteria at the end of 35-day shelf life.     

Lipoxygenase activity in wholemeal flours from Triticum monococcum, Triticum turgidum and Triticum aestivum

To minimise lipid oxidation and maintain high carotenoid and tocol concentrations in wheat flours and products, fifty-seven accessions, belonging to different Triticum species (Triticum monococcum, Triticum turgidum and Triticum aestivum), were assessed for lipoxygenase activity. The highest enzymatic activity was observed in T. aestivum (8.02 ± 0.492 μmol/min/g DM), followed by T. turgidum (3.48 ± 0.701) and by T. monococcum (0.45 ± 0.072). While the lipoxygenase was consistently high amongst T. aestivum and steadily low amongst T. monococcum samples, the T. turgidum accessions clustered in three different groups, with low (0.12-0.91 μmol/min/g DM), medium (3.10-4.17) and high (5.57-9.51) activity. Enzymatic activity was maximum in the pH range 5-6. LOX activity was higher in the germ (206 μmol/min/g DM), than in the bran (13.4) or in the endosperm (3.1).The results demonstrate that the selection of genotypes with low LOX, a factor limiting oxidative degradation, is feasible.     

Diversity of culturable microorganisms and occurrence of Listeria monocytogenes and Salmonella spp. in Tuber aestivum and Tuber melanosporum ascocarps

The aim of this study was to investigate the total mesophilic microorganisms, Pseudomonas genus, Enterobacteriaceae family, mold and yeast counts and the presence of Listeria monocytogenes and Salmonella spp on Tuber aestivum and Tuber melanosporum ascocarps. The results confirmed that the major percentage of the microorganisms, approximately 9.0 log ufc/g, were present in the peridium, the glebas of healthy truffles being practically free of microorganisms. The predominant microbial group was the Pseudomonas averaging 8.3 and 8.4 log cfu/g on T. aestivum and T. melanosporum whole ascocarps, respectively. The Enterobacteriaceae also achieved high populations, especially in T. aestivum truffles, with 6.3 log cfu/g. Molds and yeasts never exceeded 5.0 log cfu/g. The characterization of the isolates revealed that the fluorescens pseudomonads were the most prevalent. Raoultella terrigena and Enterobacter intermedius were the dominant Enterobacteriaceae. The identification of the yeast isolates revealed five species: Debaryomyces hansenii, Issatchenkia scutulata, Rhodotorula aurantiaca, Saccharomyces dairensis and Trichosporon beigelii subspecies A and B. The mold genera detected in both species of truffles were Aspergillus, Cladosporium, Penicillium and Fusarium, Trichoderma being present only in T. aestivum. L. monocytogenes was found in 10% of the samples of T. aestivum analysed but Salmonella spp. was not detected. Knowledge of the microbial population in terms of possible food borne and pathogen microorganisms is very useful for establishing successful disinfection and storage methods to prolong the shelf-life of ascocarps of T. aestivum and T. melanosporum.     

Phenolic composition of Flacourtia indica, Opuntia megacantha and Sclerocarya birrea

The phenolic acid composition of the peel and pulp of the fruits of Flacourtia indica (Burm. f.) Merr., Opuntia megacantha (L.) Mill and Sclerocarya birrea (A. Rich.) Hochst., from Zimbabwe, were analysed using traditional colorimetric as well as HPLC methods. The total phenolics, flavanoids and condensed tannin levels varied with species. Sclerocarya birrea pulp had the highest total phenolics, flavanoids and condensed tannins, i.e., 2262 μg GAE/g, 202 μg catechin/g and 6.0% condensed tannins, respectively. Flacourtia indica pulp contained the least total phenolics, flavanoids and condensed tannins 334 μg GAE/g, 41 μg catechin/g and 1.4%, respectively. There were no significant differences in the total phenolics between the peels and the pulps of the individual fruits. However, significant differences were noted in the flavanoids and the condensed tannins between the peels and pulps of the fruits assayed. Ferulic acid, caffeic acid and vanillic acid were the dominant phenolic acids in the three fruits. There were differences between the phenolic acids in the peels and the pulps of the fruits.     

Satureja horvatii essential oil: In vitro antimicrobial and antiradical properties and in situ control of Listeria monocytogenes in pork meat

The dominant compounds in Satureja horvatii oil were p-cymene (33.14%), thymol (26.11%) and thymol methyl ether (15.08%). The minimum inhibitory concentration (MIC) varied from 0.03 to 0.57 mg/mL for bacteria, and from 0.56 to 2.23 mg/mL for yeast strains, while minimum bactericidal/yeast-cidal concentration (MBC/MYC) varied from 0.07 to 1.15 mg/mL and 1.11 to 5.57 mg/mL for bacteria and yeasts, respectively. The antiradical potential of the essential oil was evaluated using hydroxyl radical (•OH) generated in Fenton reaction. The meat preserving potential of essential oil from Satureja horvatii was investigated against L. monocytogenes. Essential oil successfully inhibited development of L. monocytogenes in pork meat. Sensorial evaluation on flavor and color of meat was performed. The color and flavor of meat treated with essential oil improved after 4 days of storage. S. horvatii essential oil can act as a potent inhibitor of food spoiling microorganisms, in meat products and also can be a useful source of natural antioxidants.     

Short communication: Biofilm production characterization of mecA and mecC methicillin-resistant Staphylococcus aureus isolated from bovine milk in Great Britain

Staphylococcus aureus is an important cause of contagious intramammary infection in dairy cattle, and the ability to produce biofilm is considered to be an important virulence property in the pathogenesis of mastitis. The aim of this study was to characterize the biofilm formation capacity of methicillin-resistant Staph. aureus (MRSA), encoding mecA or mecC, isolated from bulk tank milk in Great Britain. For this purpose, 20 MRSA isolates were grown on microtiter plates to determine the biofilm production. Moreover, the spa-typing and the presence of the intercellular adhesion genes icaA and icaD were analyzed by PCR. All MRSA isolates tested belonged to 9 spa-types and were PCR-positive for the ica genes; 10 of them (50%) produced biofilm in the microtiter plate assay. This is also the first demonstration of biofilm production by mecC MRSA.     

Kinetic study of the antioxidant compounds and antioxidant capacity during germination of Vigna radiata cv. emmerald, Glycinemax cv. jutro and Glycine max cv. merit

The purpose of this study was to determine the antioxidant capacity and the content of antioxidant compounds in raw mung bean seeds and sprouts (Vigna radiata cv. emmerald) germinated for 2, 3, 4, 5 and 7 days and of soybean seeds of Glycine max cv. jutro germinated for 2, 3 and 4 days and of Glycine max cv. merit germinated for 2, 3, 4, 5 and 6 days. Antioxidant compounds, such as vitamin C and E, total phenolic compounds and reduced glutathione (GSH) were studied. Antioxidant capacity was measured by superoxide dismutase-like activity (SOD-like activity), peroxyl radical-trapping capacity (PRTC), trolox equivalent antioxidant capacity (TEAC) and inhibition of lipid peroxidation in unilamellar liposomes of egg yolk phosphatidylcholine (PC). The results indicated that changes in the contents of vitamin C, vitamin E and GSH depended on the type of legume and germination conditions. Sprouts of mung bean and soybeans provided more total phenolic compounds than did raw seeds. The SOD-like activity increased after germination of mung bean seeds for 7 days, by 308%, while no change was observed in sprouts of Glycine max cv. jutro and an increase was observed after 5 and 6 days of germination (∼20%) in Glycine max cv. merit. PRTC and TEAC increased during the germination process and retentions of 28–70% and 11–14%, respectively, for soybean, and 248% and 61%, respectively, for mung bean were observed at the end of germination. The inhibition of lipid peroxidation increased by 389% in 5–7 days’ germination of Vigna radiata cv. emmerald sprouts, and 66% in Glycine max cv. merit sprouts whilst, in Glycine max cv. jutro, germination did not cause changes in lipid peroxidation inhibition. According to the results obtained in this study, germination of mung bean and soybean seeds is a good process for obtaining functional flours with greater antioxidant capacity and more antioxidant compounds than the raw legumes.     

Simultaneous analysis of purine alkaloids and catechins in Camellia sinensis, Camellia ptilophylla and Camellia assamica var. kucha by HPLC

Simultaneous analysis of purine alkaloids and catechins in tea from dry leaves of Camellia sinensis, Camellia ptilophylla and Camellia assamica var. kucha by a reversed-phase high-performance liquid chromatography (RP-HPLC) method. This HPLC method had been proved to be appropriate for the identification and quantification of purine alkaloids and catechins, and exhibited good correlation coefficients, detection levels and recovery rates. Caffeine, theobromine, theacrine, (+)-catechin, (−)-epicatechin, (−)-epigallocatechin, (−)-epigallocatechin gallate, (−)-epicatechin gallate, (−)-gallocatechin gallate and (−)-gallocatechin were identified and quantified in the three species of genus Camellia Sect. Thea. There was 2.72% caffeine and 0.26% theobromine in C. sinensis, 4.85% theobromine in C. ptilophylla, and 1.58% theacrine, 0.94% caffeine and 0.45% theobromine in C. assamica var. kucha. Theacrine in C. sinensis and C. ptilophylla, and caffeine in C. ptilophylla were not detected. These data highlight differences in the relative proportions of purine alkaloids in the three species of Camellia Sect. Thea. In addition, different catechins were identified and quantified. The highest content of catechin in dry leaves was (−)-epigallocatechin gallate (EGCG) 3.51%, (−)-gallocatechin gallate (GCG) 9.88% and (−)-epigallocatechin gallate (EGCG) 6.78% in C. sinensis, C. ptilophylla and C. assamica var. kucha, respectively.     

Non-volatile taste components of Agaricus blazei, Agrocybe cylindracea and Boletus edulis

Three species of dried mushrooms are commercially available in Taiwan, namely Agaricus blazei (Brazilian mushroom), Agrocybe cylindracea (black popular mushroom) and Boletus edulis (king bolete), and their non-volatile taste components were studied. All mushrooms were high in contents of carbohydrate, crude fiber and protein but low in contents of crude ash and fat. Arabitol, myo-inositol, mannitol and trehalose were detected in these three mushrooms, whereas glucose was not found in B. edulis. Contents of total soluble sugars and polyols ranged from 150.33 to 225.08 mg/g. Total free amino acid contents were low in these three mushrooms and ranged from 8.97 to 14.91 mg/g. The contents of MSG-like components ranged from 1.24 to 4.40 mg/g were in the descending order of the A. blazei, A. cylindracea and B. edulis. Total 5′-nucleotides contents of A. blazei and A. cylindracea were higher than that of B. edulis whereas flavor 5′-nucleotides content of A. blazei was higher than those of A. cylindracea and B. edulis. Equivalent umami concentrations values in three mushrooms ranged from 10.46 to 135.90 g per 100 g. Overall, these three mushrooms possessed highly umami taste.     

The in vitro and in vivo effects of yerba mate (Ilex paraguariensis) extract on adipogenesis

The aim of this study was to evaluate the effects of yerba mate extract and its principal bioactive compounds on adipogenesis. The anti-adipogenic effects of yerba mate, chlorogenic acid, quercetin and rutin were evaluated in 3T3-L1 cells using a PCR array. The results obtained in vitro were validated in vivo in a high-fat diet-induced model of obesity. The in vitro and in vivo results demonstrated that yerba mate extract down-regulated the expression of genes that regulate adipogenesis, such as Creb-1and C/EBPα, and the extract up-regulated the expression of genes related to the inhibition of adipogenesis, including Dlk1, Gata2, Gata3, Klf2, Lrp5, Pparγ₂, Sfrp1, Tcf7l2, Wnt10b, and Wnt3a. In summary, it was demonstrated that yerba mate and its bioactive compounds regulate the expression of genes related to in vitro adipogenesis. Furthermore, yerba mate might regulate adipogenesis through the Wnt pathway.     

Antioxidant properties of Agaricus blazei, Agrocybe cylindracea, and Boletus edulis

Three mushrooms are currently available in Taiwan, including Agaricus blazei, Agrocybe cylindracea, and Boletus edulis. Their ethanolic and hot water extracts were prepared and antioxidant properties studied. Ethanolic extracts from three mushrooms were more effective than hot water extracts in antioxidant activity using the conjugated diene method and scavenging ability on 1,1-diphenyl-2-picrylhydrazyl radicals whereas hot water extracts were more effective in reducing power, scavenging ability on hydroxyl radials and chelating ability on ferrous ions as evidenced by their lower EC₅₀ values. Overall, for both extracts, B. edulis was more effective among antioxidant properties assayed. Naturally occurring antioxidant components including total tocopherols (3.18-6.18 mg/g) and total phenols (5.67-5.81 mg/g) were found in the extracts and their contents were associated (r=0.636-0.907) with EC₅₀ value of antioxidant properties. Based on the results obtained, both extracts from these three mushrooms were effective in antioxidant properties.     

Citrus paradisi and Citrus sinensis flavonoids: Their influence in the defence mechanism against Penicillium digitatum

Citrus peel is rich in flavanone glycosides and polymethoxyflavones. In view of their importance for industrial application as well as for their pharmacological properties, their content was analyzed in the mature fruits of several Citrus paradisi (grapefruit) and Citrus sinensis (orange) varieties, with a view to select the most interesting for isolation. The results shows that the Star Ruby grapefruit and the Sanguinelli orange stand out for their high contents of naringin and hesperidin, respectively. The presence of the polymethoxyflavones nobiletin, heptamethoxyflavone and tangeretin, could be ascertained in all the grapefruit varieties analysed. Higher polymethoxyflavone levels were recorded in orange, with Valencia Late showing the greatest nobiletin, sinensetin and tangeretin contents and Navelate the highest heptamethoxyflavone levels. An in vitro study revealed that these compounds acted as antifungal agents against Penicillium digitatum, the polymethoxyflavones being more active than the flavanones in this respect. The possible participation of these phenolic compounds in the defence mechanism of Citrus against P. digitatum is discussed.     

Cloning and Characterization of clpB in Acetobacter pasteurianus NBRC 3283

The clpB gene in Acetobacter pasteurianus was cloned and characterized. Although the clpB gene was transcribed in response to a temperature shift and exposure to ethanol, the clpB disruption mutant was only affected by high temperature, suggesting that the ClpB protein is closely associated with heat resistance in A. pasteurianus.     

Short-term toxicological evaluation of Terminalia catappa, Pentaclethra macrophylla and Calophyllum inophyllum seed oils in rats

The purpose of this study was to evaluate the toxicological effects of feeding the oils of Calophyllum inophyllum, Pentaclethra macrophylla and Terminalia catappa to rats. The effects on physical appearance, feed intake, weight gain, plasma and tissue cholesterol and triacyglycerol levels in rats with 5% of the oils in normal rat feed were determined. Weekly monitoring of the rats showed good physical appearance and steady weight gain, with no mortality recorded for the period of the study. Haematological analysis of the rats indicated that they were not anaemic. Histopathotogical examination of the sections of the heart, liver, kidney and spleen revealed moderate (T. catappa oil) to severe fatty change and necrosis in the liver. Glomerulonephrotic changes in the kidneys of rats fed with T. catappa oil were moderate, while it was severe in the group fed with P. macrophylla oil. Severe myocardiac necrosis as well as atherosclerotic clefts in vasa vasori was observed in the vasa vasori of the hearts of rats fed with P. macrophylla oil. This change was moderate in the heart of rats fed with C. inophyllum, while no such observation was made in the group fed with T. catappa oil. There was a significant difference in the plasma cholesterol levels of the rats fed with C. inophyllum and T. catappa oils when compared with the control rats, while those fed with P. macrophylla oil had no significant difference. The oil of T. catappa appears more suitable for consumption than the oils from C. inophyllum and P. macrophylla. Fatty acid analysis of the oils showed that they have high amounts of unsaturated fatty acids with linoleic and oleic acids as the major ones.     

Distribution of the Medea factor M in populations of Tribolium castaneum (Herbst) in the United States

The distribution of the maternally acting, selfish gene Medea⁴ (M⁴) was determined in populations of the red flour beetle, Tribolium castaneum (Herbst), collected in the southern and midwestern United States. We found clear evidence for the existence of two major regional subpopulations, with a boundary that roughly corresponds to 33°N latitude. All 26 strains collected in 10 states north of this latitude were homozygous for the M⁴ allele, while only two of 29 strains collected in six states south of this latitude were homozygous for the allele. Of the remaining 27 southern strains, 21 lacked the M⁴ allele entirely, while six contained a mixture of M⁴ and non-M⁴ alleles. This is the first evidence of either the existence of biotypes or the presence of major barriers to gene flow in wild populations of this ubiquitous insect species.     

Application of Kluyveromyces marxianus, Lactobacillus delbrueckii ssp. bulgaricus and L. helveticus for sourdough bread making

The application of Kluyveromyces marxianus (IFO 288), Lactobacillus delbrueckii ssp. bulgaricus (ATCC 11842) and Lactobacillus helveticus (ATCC 15009) as starter cultures for sourdough bread making was examined. Production of lactic and acetic acids, bread rising, volatile composition, shelf-life and organoleptic quality of the sourdough breads were evaluated. The amount of starter culture added to the flour, the dough fermentation temperature and the amount of sourdough used were examined in order to optimise the bread making process. The use of mixed cultures led to higher total titratable acidities and lactic acid concentrations compared to traditionally made breads. Highest acidity (3.41 g lactic acid/kg of bread) and highest resistance to mould spoilage were observed when bread was made using 50% sourdough containing 1% K. marxianus and 4% L. delbrueckii ssp. bulgaricus. The use of these cultures also improved the aroma of sourdough breads, as shown by sensory evaluations and as revealed by GC–MS analysis.     

Phosphine-induced sterility in Ephestia cautella (Lepidoptera: Pyralidae)

The present study was conducted to explore the sterilizing effect of phosphine on Ephestia cautella. Three to four-day old pupae were exposed to 0, 0.007, 0.014, 0.021, 0.028, 0.035, 0.042 and 0.049 mg/l of phosphine for 24 h at room temperature. The normally developed adults of each treatment were pair mated and several physiological and genetical parameters were investigated. The results showed gradual increases with dose in pupal mortality and adult malformation. The crossing results showed no changes in adult longevity and mating frequency. However, consistent reduction was noted in the fecundity and fertility of normally emerged adults. Preliminary data indicated similar levels of inherited sterility in the F₁ adults raised from the above mentioned crosses.