Showing papers in "Journal of Applied Microbiology in 2015"

The use of probiotics in aquaculture

Abstract: This study aims to present comprehensive notes for the use of probiotics in aquaculture. Probiotics have been proven to be positive promoters of aquatic animal growth, survival and health. In aquaculture, intestines, gills, the skin mucus of aquatic animals, and habitats or even culture collections and commercial products, can be sources for acquiring appropriate probiotics, which have been identified as bacteria (Gram-positive and Gram-negative) and nonbacteria (bacteriophages, microalgae and yeasts). While a bacterium is a pathogen to one aquatic animal, it can bring benefits to another fish species; a screening process plays a significant role in making a probiotic species specific. The administration of probiotics varies from oral/water routine to feed additives, of which the latter is commonly used in aquaculture. Probiotic applications can be either mono or multiple strains, or even in combination with prebiotic, immunostimulants such as synbiotics and synbiotism, and in live or dead forms. Encapsulating probiotics with live feed is a suitable approach to convey probiotics to aquatic animals. Dosage and duration of time are significant factors in providing desired results. Several modes of actions of probiotics are presented, while some others are not fully understood. Suggestions for further studies on the effects of probiotics in aquaculture are proposed.

Phospholipid fatty acid profiling of microbial communities a review of interpretations and recent applications

Abstract: Summary Profiling of microbial communities in environmental samples often utilizes phospholipid fatty acid (PLFA) analysis. This method has been used for more than 35 years and is still popular as a means to characterize microbial communities in a diverse range of environmental matrices. This review examines the various recent applications of PLFA analysis in environmental studies with specific reference to the interpretation of the PLFA results. It is evident that interpretations of PLFA results do not always correlate between different investigations. These discrepancies in interpretation and their subsequent applications to environmental studies are discussed. However, in spite of limitations to the manner in which PLFA data are applied, the approach remains one with great potential for improving our understanding of the relationship between microbial populations and the environment. This review highlights the caveats and provides suggestions towards the practicable application of PLFA data interpretation.

Lipid production by yeasts growing on biodiesel‐derived crude glycerol: strain selection and impact of substrate concentration on the fermentation efficiency

Abstract: Aims To screen yeasts in relation to the potential to produce single cell oil (SCO) from biodiesel-derived glycerol and to enhance SCO production in Lipomyces starkeyi and Rhodosporidium toruloides yeasts. Methods and results Yarrowia lipolytica, Cryptococcus curvatus, R. toruloides and L. starkeyi were grown in nitrogen-limited flask cultures. Yarrowia lipolytica strains produced citric acid and mannitol. Lipomyces starkeyi DSM 70296 and R. toruloides NRRL Y-27012 showed potential for SCO production, and were cultivated at increasing the initial glycerol concentrations with the initial nitrogen concentration remaining constant. Significant biomass and SCO production were reported even in cultures with high initial glycerol concentrations (i.e. 180 g l(-1) ). Lipid quantities of c. 12 g l(-1) (lipid in dry cell weight 35-40%) were obtained for both L. starkeyi and R. toruloides, quite high values compared with literature values for oleaginous micro-organisms growing on glycerol. However, these strains exhibited different kinetic profiles in the synthesis of intracellular polysaccharides. Lipomyces starkeyi produced a significant quantity of polysaccharides (c. 7 g l(-1) ). The yeast lipids contained mainly oleic and palmitic and to a lesser extent linoleic and stearic acids. Conclusions Lipomyces starkeyi and R. toruloides are potential SCO producers from crude glycerol. Significance and impact of the study Very scarce numbers of reports have indicated the production of SCO by L. starkeyi and R. toruloides growing on glycerol. We report here that these yeasts are able efficiently to convert raw glycerol into SCO, while L. starkeyi also synthesizes intracellular polysaccharides in marked quantities.

Assessment of human virus removal during municipal wastewater treatment in Edmonton, Canada

Abstract: Aims To assess the removal of viruses through the multiple steps of wastewater treatment in a full-scale municipal wastewater treatment plant in Alberta, Canada. Methods and Results Samples were collected after each of the five treatment steps for a period of 16 months. The amount of viruses and their infectivity were analysed using real-time quantitative PCR (qPCR) and integrated viral cell culture (ICC), respectively. Bacterial indicator Escherichia coli was also tested using membrane filtration. Seven viruses including Norovirus (NoV), Rotavirus (RV), Sapovirus (SaV), Astrovirus (AsV), Adenovirus (AdV), Enterovirus (EV) and JC virus (JCV) were detected in 16 primary effluents in which infectious viruses were present. Different treatment steps showed various efficiencies in virus removal, with membrane ultrafiltration as the most effective at 4·6–7·0 log reduction. Conclusions We observed high prevalence of viruses in raw wastewater and different viral reduction after various treatment steps. The discharge of treated wastewater with infectious viruses represents potential risks to human, animal and environmental health. Significance and Impact of the Study This study provides a comprehensive assessment of the removal of NoV, RV, SaV, AsV, AdV, EV, JCV and Reovirus from wastewater by current procedures of municipal wastewater treatment and discusses the applicability of various viruses as viral indicators for water quality.

Putative bacterial volatile-mediated growth in soybean (Glycine max L. Merrill) and expression of induced proteins under salt stress

Abstract: Aims Plant root-associated rhizobacteria elicit plant immunity referred to as induced systemic tolerance (IST) against multiple abiotic stresses. Among multibacterial determinants involved in IST, the induction of IST and promotion of growth by putative bacterial volatile compounds (VOCs) is reported in the present study. Methods and Results To characterize plant proteins induced by putative bacterial VOCs, proteomic analysis was performed by MALDI-MS/MS after exposure of soybean seedlings to a new strain of plant growth promoting rhizobacteria (PGPR) Pseudomonas simiae strain AU. Furthermore, expression analysis by Western blotting confirmed that the vegetative storage protein (VSP), gamma-glutamyl hydrolase (GGH) and RuBisCo large chain proteins were significantly up-regulated by the exposure to AU strain and played a major role in IST. VSP has preponderant roles in N accumulation and mobilization, acid phosphatase activity and Na+ homeostasis to sustain plant growth under stress condition. More interestingly, plant exposure to the bacterial strain significantly reduced Na+ and enhanced K+ and P content in root of soybean seedlings under salt stress. In addition, high accumulation of proline and chlorophyll content also provided evidence of protection against osmotic stress during the elicitation of IST by bacterial exposure. Conclusions The present study reported for the first time that Ps. simiae produces a putative volatile blend that can enhance soybean seedling growth and elicit IST against 100 mmol l−1 NaCl stress condition. Significance and Impact of the Study The identification of such differentially expressed proteins provide new targets for future studies that will allow assessment of their physiological roles and significance in the response of glycophytes to stresses. Further work should uncover more about the chemical side of VOC compounds and a detailed study about their molecular mechanism responsible for plant growth.

Antifungal modes of action of tea tree oil and its two characteristic components against Botrytis cinerea

Abstract: Aims The essential oil of Melaleuca alternifolia (tea tree) has been evaluated as a potential eco-friendly antifungal agent against Botrytis cinerea. In this study, we investigated the antifungal activity and mode of action of tea tree oil (TTO) and its components against B. cinerea. Methods and Results Of the components we tested in contact phase, terpinen-4-ol had the highest antifungal activity, followed by TTO, α-terpineol, terpinolene, then 1,8-cineole. As one of characteristic components of TTO, terpinen-4-ol treatment led to pronounced alterations in mycelial morphology, cellular ultrastructure, membrane permeability under scanning electron microscope, transmission electron microscope and fluorescent microscope, and also reduced the ergosterol content of fungi. As another characteristic component, 1,8-cineole caused serious intracellular damage but only slightly affected B. cinerea otherwise. When terpinen-4-ol and 1,8-cineole were used together, the synergistic antifungal activity was significantly higher than either component by itself. Conclusions The results of our study confirmed that terpinen-4-ol and 1,8-cineole act mainly on the cell membranes and organelles of B. cinerea, respectively, and when combined are similar to TTO in antifungal activity due to their differences. Significance and Impact of the Study Understanding the mechanism of terpinen-4-ol and 1,8-cineole antifungal action to B. cinerea is helpful for investigation on their synergistic effect and explaining antifungal action modes of TTO.

Antibiofilm potential of flavonoids extracted from Moringa oleifera seed coat against Staphylococcus aureus, Pseudomonas aeruginosa and Candida albicans.

Abstract: Aims The increased microbial drug resistance due to biofilms and the side effects associated with the use of conventional drugs is still a major concern in the medical fraternity. This work evaluates the antibiofilm potential of flavonoids extracted from Moringa oleifera seed coat (SC) in search for green and effective alternatives for overcoming menace of biofilms. Methods and Results The study evaluated the minimum inhibitory concentration (MIC) of flavonoids against respective test organisms, inhibition of initial cell attachment as well as disruption of preformed biofilms and metabolic activity of treated biofilms. Mutagenicity and cytotoxicity as well as characterization of the active component were also carried out. Although Pseudomonas aeruginosa showed the lowest MIC of 0·05 mg ml−1, the action of flavonoids and gentamicin on initial cell attachment revealed a comparable effect against bacterial biofilms, i.e. Staphylococcus aureus and Pseudomonas aeruginosa with approx. 80% inhibition compared to Candida albicans. Disruption of the preformed biofilms revealed that susceptibility of P. aeruginosa began as early as 4 h of exposure to flavonoids with 88% growth inhibition at the end of 24-h incubation. Encouragingly, t-test analysis on the effect of the extract and the standard antibiotic against each organism indicated no significant variance at P < 0·05. A drastic low metabolic activity exhibited by the treated biofilms as compared to the untreated ones was further supportive of the antibiofilm potential of seed coat flavonoids. Conclusion The bioactive component from M. oleifera seed coat has exhibited antibiofilm potential against the test organisms belonging to Gram positive, Gram negative and yeast. Significance and Impact of the Study Antibiofilm potential and biosafety of plant-based flavonoids from M. oleifera seed coat reveal a prospective active principle that could be of use in biofilm-associated menace.

Probiotic potentials of yeasts isolated from some cereal-based Nigerian traditional fermented food products.

Abstract: Aims To determine the starter culture and multifunctional potentials of yeast strains from some cereal-based Nigerian traditional fermented food products. Methods and Results Yeast isolates were screened for enzyme production and identified by sequencing the D1/D2 region of 26S rDNA. Pichia kluyveri LKC17, Issatchenkia orientalis OSL11, Pichia kudriavzevii OG32, Pichia kudriavzevii ROM11 and Candida tropicalis BOM21 exhibited the highest protease, lipase and phytase activity. They were selected and further evaluated for gastrointestinal survival and adherence ability. Although strain-specific, they retained viability at 37°C and showed survival at pH 2·0., I. orientalis OSL11 showed the highest survival at 2% bile salts concentration and P. kudriavzevii ROM11 showed the least survival. The yeast strains showed strong autoaggregation ability (81·24–91·85%) and hydrophobicity to n-hexadecane (33·61–42·30%). The highest co-aggregation ability was detected for P. kudriavzevii OG32 and Escherichia coli (71·57%). All the yeast strains removed cholesterol in the range of 49·03–74·05% over 48 h and scavenged for free radicals in methanol reaction system. Conclusions In this study, we isolated new yeast strains with multifunctional potentials that can be used as functional starter cultures to produce cereal-based probiotic products. Significance and Impact of the Study The development of probiotic yeast strains as starter culture to improve the quality attributes and confer functional value on cereal-based traditional fermented foods is beneficial.

Whole cell kinetics of ureolysis by Sporosarcina pasteurii.

Abstract: NSF's Collaborations in Mathematical Geosciences (CMG) program no. DMS-0934696; DOE Subsurface Biogeochemical Research (SBR) Program no. DE-FG02-09ER64758; University Scholars Program (USP);U.S. DOE DE-FE0004478, DE-FE0009599 and DE-FG02-13ER86571

Thermophilic fungi as new sources for production of cellulases and xylanases with potential use in sugarcane bagasse saccharification.

Abstract: AIMS To obtain new cellulases and xylanases from thermophilic fungi; evaluate their potential for sugarcane bagasse saccharification. METHODS AND RESULTS Thirty-two heat-tolerant fungi were isolated from the environment, identified (morphological/molecular tools) and the production of the enzymes was evaluated by solid state fermentation using lignocellulosic materials as substrates. Myceliophthora thermophila JCP 1-4 was the best producer of endoglucanase (357·51 U g(-1) ), β-glucosidase (45·42 U g(-1) ), xylanase (931·11 U g(-1) ) and avicelase (3·58 U g(-1) ). These enzymes were most active at 55-70°C and stable at 30-60°C. Using crude enzymatic extract from M. thermophila JCP 1-4 to saccharify sugarcane bagasse pretreated with microwaves and glycerol, glucose and xylose yields obtained were 15·6 and 35·13% (2·2 and 1·95 g l(-1) ), respectively. CONCLUSIONS All isolated fungi have potential to produce the enzymes; M. thermophila JCP 1-4 enzymatic extract have potential to be better explored in saccharification experiments. Pretreatment improved enzymatic saccharification, as sugar yields were much higher than those obtained from in natura bagasse. SIGNIFICANCE AND IMPACT OF THE STUDY Myceliophthora thermophila JCP 1-4 produces avicelase (not commonly found among fungi; important to hydrolyse crystalline cellulose) and a β-glucosidase resistant to glucose inhibition, interesting characteristics for saccharification experiments.

Soil Fungi for Mycoremediation of Arsenic Pollution in Agriculture Soils

Abstract: Aims Soil arsenic (As) contamination of food-chains and public health can be mitigated through fungal bioremediation. To enumerate culturable soil fungi, soils were collected from the As-contaminated paddy fields (3–35 mg kg−1) of the middle Indo-Gangetic Plains. Methods and Results Total 54 fungal strains were obtained and identified at their molecular level. All strains were tested for As tolerance (from 100 to 10 000 mg l−1 arsenate). Fifteen fungal strains, tolerant to 10 000 mg l−1 arsenate, were studied for As removal in-vivo for 21 days by cultivating them individually in potato dextrose broth enriched with 10 mg l−1 As. The bioaccumulation of As in fungal biomass ranged from 0·023 to 0·259 g kg−1. The biovolatilized As ranged from 0·23 to 6·4 mg kg−1. Conclusions Higher As bioaccumulation and biovolatilization observed in the seven fungal strains, Aspergillus oryzae FNBR_L35; Fusarium sp. FNBR_B7, FNBR_LK5 and FNBR_B3; Aspergillus nidulans FNBR_LK1; Rhizomucor variabilis sp. FNBR_B9; and Emericella sp. FNBR_BA5. These fungal strains were also tested and found suitable for significant plant growth promotion in the calendula, withania and oat plants in a greenhouse based pot experiment. Significance and Impact of Study These fungal strains can be used for As remediation in As-contaminated agricultural soils.

Probiotic Lactobacillus rhamnosus GR‐1 and Lactobacillus reuteri RC‐14 exhibit strong antifungal effects against vulvovaginal candidiasis‐causing Candida glabrata isolates

Abstract: Aims: This study investigates the antagonistic effects of the probiotic strains Lactobacillus rhamnosus GR-1 and Lactobacillus reuteri RC-14 against vulvovaginal candidiasis (VVC)-causing Candida glabrata. Methods and Results: Growth inhibitory activities of Lact. rhamnosus GR-1 and Lact. reuteri RC-14 strains against C. glabrata were demonstrated using a spot overlay assay and a plate-based microtitre assay. In addition, these probiotic lactobacilli strains also exhibited potent candidacidal activity against C. glabrata, as demonstrated by a LIVE/DEAD yeast viability assay performed using confocal laser scanning microscopy. The metabolic activities of all C. glabrata strains were completely shut down in response to the challenges by the probiotic lactobacilli strains. In addition, both probiotic lactobacilli strains exhibited strong autoaggregation and coaggregation phenotypes in the presence of C. glabrata, which indicate that these lactobacilli strains may exert their probiotic effects through the formation of aggregates and, thus the consequent prevention of colonization by C. glabrata. Conclusions: Probiotic Lact. rhamnosus GR-1 and Lact. reuteri RC-14 strains exhibited potent antagonistic activities against all of the tested C. glabrata strains. These lactobacilli exhibited antifungal effects, including those attributed to their aggregation abilities, and their presence caused the cessation of growth and eventual cell death of C. glabrata. Significance and Impact of the Study: This is the first study to report on the antagonistic effects of these probiotic lactobacilli strains against the non-Candida albicans Candida (NCAC) species C. glabrata.

Inhibition of Candida albicans adhesion on medical‐grade silicone by a Lactobacillus‐derived biosurfactant

Abstract: Aims The study aimed at investigating the ability of biosurfactant (BS) produced by a Lactobacillus brevis isolate (CV8LAC) to inhibit adhesion and biofilm formation of Candida albicans on medical-grade silicone elastomeric disks (SEDs). Methods and Results Biosurfactant activity was evaluated at physiological conditions, by means of co-incubation and precoating assays. Additionally, BS extract was tested for antifungal susceptibility against C. albicans in both planktonic and sessile form. Biofilm covered surface and hyphae and blastospores occurrence were quantified by scanning electron microscopy (SEM) and image analysis. BS did not inhibit growth of C. albicans in both planktonic and sessile form. Nevertheless, co-incubation with 2000 μg ml−1 BS significantly reduced biofilm formation on SEDs surface by 89, 90 and 90% after 24, 48 and 72 h of incubation. Fungal adhesion and biofilm formation to precoated SEDs was reduced by 62, 53, 50 and 44% after 1·5, 24, 48 and 72 h. SEM showed a significant reduction of biofilm covered surface in precoated disks but no differences in the production of hyphae or blastospores, except at 1·5 h of incubation. Conclusions This study demonstrated that CV8LAC BS has the ability to counteract significantly the initial deposition of C. albicans to silicone surfaces and to effectively slow biofilm growth. Significance and Impact of the Study The anti-adhesive properties of the CV8LAC BS suggest a potential role of the coating for preventing fungal infection associated to silicone medical devices.

Diversity of food-borne Bacillus volatile compounds and influence on fungal growth

Abstract: Aims To evaluate the antifungal activity of the volatile organic compounds (VOCs) produced by 75 different food-borne Bacillus species against Aspergillus niger, Aspergillus flavus, Aspergillus parasiticus, Aspergillus clavatus, Fusarium oxysporum f. sp. lactucae and Moniliophthora perniciosa and to determine the VOCs responsible for the inhibition. Methods and Results Bacillus strains inhibited fungal growth, although with different inhibition grades, with Bacillus subtilis, Bacillus amyloliquefaciens and Bacillus cereus strains as the best antifungal VOCs producers. While M. perniciosa DM4B and F. oxysporum f.sp. lactucae MA28 were the most sensitive fungi, A. parasiticus MG51 showed the greatest resistance to Bacillus VOCs exposure. Thirty-seven compounds were detected by SPME-GC-MS analysis, although similar patterns in volatile compounds were evidenced within the species, interspecific VOCs differences determined different effects on fungal growth. Multiple partial least regression (MPLRS) and antifungal activity of the individual VOCs revealed that only propanone, 1-butanol, 3-methyl-1-butanol, acetic acid, 2-methylpropanoic acid, carbon disulphide, 3-methylbutanoic acid and ethyl acetate were responsible for mycelia inhibition of M. perniciosa DM4B and F. oxysporum f.sp. lactucae MA28. Conclusions The antagonistic activity of the Bacillus VOCs was demonstrated, although it cannot easily be explained through the action of a single molecule, thus a holistic approach could be more appropriate to estimate the fungal growth inhibition. Significance and Impact of the Study VOCs produced by Bacillus from cooked food can be considered as promising antifungal compounds useful in the control of fungal plant pathogens. This study investigates for the first time the correlation between mycelia inhibition of M. perniciosa and F. oxysporum f. sp. lactucae and the VOCs emitted by the Bacillus species.

A review: what is the spermosphere and how can it be studied?

Abstract: The spermosphere is the zone surrounding seeds where interactions between the soil, microbial communities and germinating seeds take place. The concept of the spermosphere is usually only applied during germination sensu stricto. Despite the transient nature of this very small zone of soil around the germinating seed, the microbial activities which occur there may have long-lasting impacts on plants. The spermosphere is indirectly characterized by either (i) seed exudates, which could be inhibitors or stimulators of micro-organism growth or (ii) the composition of the microbiome on and around the germinating seeds. The microbial communities present in the spermosphere directly reflect that of the germination medium or are host-dependent and influenced quantitatively and qualitatively by host exudates. Despite its strong impact on the future development of plants, the spermosphere remains little studied. This can be explained by the technical difficulties related to characterizing this concept due to its short duration, small size and biomass, and the number and complexity of the interactions that take place. However, recent technical methods, such as metabolite profiling, combining phenotypic methods with DNA- and RNA-based methods, could be used to investigate seed exudates, microbial communities and their interactions with the soil environment.

The state of autotrophic ethanol production in Cyanobacteria

Abstract: Ethanol production directly from CO2 , utilizing genetically engineered photosynthetic cyanobacteria as a biocatalyst, offers significant potential as a renewable and sustainable source of biofuel. Despite the current absence of a commercially successful production system, significant resources have been deployed to realize this goal. Utilizing the pyruvate decarboxylase from Zymomonas species, metabolically derived pyruvate can be converted to ethanol. This review of both peer-reviewed and patent literature focuses on the genetic modifications utilized for metabolic engineering and the resultant effect on ethanol yield. Gene dosage, induced expression and cassette optimizat-ion have been analyzed to optimize production, with production rates of 0·1-0·5 g L(-1) day(-1) being achieved. The current 'toolbox' of molecular manipulations and future directions focusing on applicability, addressing the primary challenges facing commercialization of cyanobacterial technologies are discussed.

Autochthonous starter cultures and indigenous grape variety for regional wine production.

Abstract: Aims To characterize Oenococcus oeni strains isolated from North-Apulian wines where malic acid degradation is usually achieved by spontaneous fermentations, and to determine the influence of bacterial inoculation time on the malolactic performances in ‘Nero di Troia’ wine using a complete autochthonous microbial regime. Methods and Results Oenococcus oeni strains from wines produced with the autochthonous (Apulia Region, southern Italy) grape variety ‘Uva di Troia’ were isolated, selected and characterized. Multilocus sequence typing and variable number tandem repeat analysis were used to investigate intraspecific diversity. Oenococcus oeni strains were tested in co-inoculation and in sequential inoculation, with two autochthonous yeast strains previously isolated from ‘Nero di Troia’ wine. After a preliminary screening using co-inoculation regime, the O. oeni strains were grouped in reason of the different behaviour in malic acid performances. Results suggested that the efficient degradation of malic acid in co-inoculation is a strain-dependent characteristic. Conclusions Autochthonous yeast/bacterium combinations were identified as starter culture, and used in a co-inoculation approach, for vinification of regional wines. Significance and Impact of the Study The ‘microbial terroir’ of typical fermented food and beverage production represents a dynamic sector of applied research in food microbiology. In this work, we propose the use of autochthonous bacteria and yeast for wine production from an indigenous grape variety.

Antimicrobial activity of hop extracts against foodborne pathogens for meat applications

Abstract: Aims The objective of this study was the fundamental investigation of the antimicrobial efficiency of various hop extracts against selected foodborne pathogens in vitro, as well as their activity against Listeria monocytogenes in a model meat marinade and on marinated pork tenderloins. Methods and Results In a first step, the minimum inhibitory concentrations (MIC) of three hop extracts containing either α- or β-acids or xanthohumol were determined against test bacteria including L. monocytogenes, Staphylococcus aureus, Salmonella enterica and Escherichia coli by a colorimetric method based on the measurement of bacterial metabolic activity. Moreover, the influence of either lactic or citric acid on the antimicrobial activity of the hop extracts was evaluated. The efficiency of hop extracts as a natural food preservative was then tested in a model meat marinade at 2 and 8°C, respectively, and finally on marinated pork. The experiments showed that Gram-positive bacteria were strongly inhibited by hop extracts containing β-acids and xanthohumol (MIC values of 6·3 and 12·5 ppm, respectively), whereas the antimicrobial activity of the investigated α-acid extract was significantly lower (MIC values of 200 ppm). Gram-negative bacteria were highly resistant against all tested hop extracts. Acidification of the test media led to a decrease of the MIC values. The inhibitory activity of the hop extracts against L. monocytogenes was strongly reduced in a fat-containing model meat marinade, but the efficiency of β-acids in this matrix could be increased by lowering pH and storage temperatures. By applying 0·5 % β-acids at pH = 5 in a model marinade, the total aerobic count of pork tenderloins was reduced up to 0·9 log10 compared with marinated pork without hop extract after 2 weeks of storage at 5°C. Conclusions β-acid containing hop extracts have proven to possess a high antimicrobial activity against Gram-positive bacteria in vitro and in a practice-related application for food preservation. Significance and Impact of the Study Antimicrobial hop extracts could be used as natural preservatives in food applications to extend the shelf life and to increase the safety of fresh products.

Antifungal activity of various essential oils against Rhizoctonia solani and Macrophomina phaseolina as major bean pathogens

Abstract: Aims The main objective of this study was to investigate the effect of various essential oils (EOs) to decrease the activity of cell wall degrading enzymes (CWDEs) produced by fungal phytopathogens, which are associated with disease progress. Also, effect of seed treatment and foliar application of peppermint EO and its main constituent, menthol, on diseases caused by two necrotrophic pathogens on bean was investigated. Methods and Results Antifungal activity of EOs on Rhizoctonia solani and Macrophomina phaseolina, as bean pathogens, was evaluated. The EOs of Mentha piperita, Bunium persicum and Thymus vulgaris revealed the highest antifungal activity against fungi. The EO of M. piperita had the lowest minimum inhibitory concentration (MIC) for R. solani among the three EOs tested. This pathogen did not grow in the presence of M. piperita, B. persicum and T. vulgaris EOs at 850, 1200 and 1100 ppm concentrations, respectively. The B. persicum EO had the lowest MIC for M. phaseolina as this fungus did not grow in the presence of M. piperita, B. persicum and T. vulgaris EOs at concentrations of 975, 950 and 1150 ppm, respectively. Hyphae exposed to EOs showed structural changes. Activities of cellulase and pectinase, as main CWDEs of pathogens, decreased by EOs at low concentration without effect on fungal growth. Seed treatment and foliar application of peppermint EO and/or menthol significantly reduced the development of bean diseases caused by both fungi. Higher capability of menthol than peppermint EO in decreasing diseases on bean was observed. Conclusions Reducing CDWEs activity is a mechanism of EOs' effect on fungi. Higher antifungal activity of menthol compared to peppermint EO was observed not only in vitro but also in vivo. Significance and Impact of the Study Effect of EOs on CWDEs involved in pathogenesis is described in this study for the first time. Menthol can be used as a botanical fungicide to control destructive fungal diseases on bean.

In vitro evaluation of the acquisition of resistance, antifungal activity and synergism of Brazilian red propolis with antifungal drugs on Candida spp.

Abstract: Aims To evaluate the ability of Candida parapsilosis and Candida glabrata to develop phenotypic resistance to a benzophenone enriched fraction obtained from Brazilian red propolis (BZP-BRP) as compared to fluconazole (FLC). To investigate possible synergy between BZP-BRP and FLC and anidulafungin (AND). Methods and Results To analyse the development of resistance, isolates susceptible to these antifungals were cultured in increasing concentrations of FLC and BZP-BRP. The increase in FLC minimum inhibitory concentration for all isolates was evident and the majority developed resistance, whereas none isolated became less susceptible to BZP-BRP. Synergism was investigated by checkerboard method. BZP-BRP demonstrated synergy with FLC and indifference with AND for most isolates. Conclusions In conclusion, the synergism observed with FLC suggests that BZP-BRP could be a possible therapeutic strategy for the treatment of infections related to FLC-resistant Candida sp. Significance and Impact of the Study The indiscriminate use of antifungals results in the emergence of drug-resistant strains among previously susceptible populations. BZP-BRP can become an alternative for the treatment of persistent infections caused by Candida sp.

Review of established and innovative detection methods for carbapenemase-producing Gram-negative bacteria.

Abstract: The minimal antibiotic options for carbapenemase-producing Gram-negative bacteria necessitate their rapid detection. A literature review of a variety of phenotypic and genotypic methods is presented. Advances in culture methods and screening media are still subject to long incubation hours. Biochemical methods have shorter turnaround times and higher sensitivities and specificities, but cannot differentiate between various types and variants. Spectrophotometric methods are cheap and efficient, but are uncommon in many clinical settings, while the MALDI-TOF MS is promising for species identification, typing and resistance gene determination. Although next generation sequencing (NGS) technologies provide a better platform to detect, type and characterize carbapenem-resistant bacteria, the different NGS platforms, the large computer memories and space needed to process and store genomic data and the nonuniformity in data analysis platforms are still a challenge. The sensitivities, specificities and turnaround times recorded in the various studies reviewed favours the use of the biochemical tests (Carba NP or Rapid Carb screen tests) for the detection of putative carbapenemase-producing isolates. MALDI-TOF MS and/or molecular methods like microarray, loop-mediated isothermal amplification and real-time multiplex PCR assays could be used for further characterization in a reference laboratory. NGS may be used for advanced epidemiological and molecular studies.

Production of arabitol by yeasts: current status and future prospects

Abstract: Arabitol belongs to the pentitol family and is used in the food industry as a sweetener and in the production of human therapeutics as an anticariogenic agent and an adipose tissue reducer. It can also be utilized as a substrate for chemical products such as arabinoic and xylonic acids, propylene, ethylene glycol, xylitol and others. It is included on the list of 12 building block C3-C6 compounds, designated for further biotechnological research. This polyol can be produced by yeasts in the processes of bioconversion or biotransformation of waste materials from agriculture, the forest industry (l-arabinose, glucose) and the biodiesel industry (glycerol). The present review discusses research on native yeasts from the genera Candida, Pichia, Debaryomyces and Zygosaccharomyces as well as genetically modified strains of Saccharomyces cerevisiae which are able to utilize biomass hydrolysates to effectively produce L- or D-arabitol. The metabolic pathways of these yeasts leading from sugars and glycerol to arabitol are presented. Although the number of reports concerning microbial production of arabitol is rather limited, the research on this topic has been growing for the last several years, with researchers looking for new micro-organisms, substrates and technologies.

Deinococcus as new chassis for industrial biotechnology: biology, physiology and tools

Abstract: Deinococcus spp are among the most radiation-resistant micro-organisms that have been discovered. They show remarkable resistance to a range of damage caused by ionizing radiation, desiccation, UV radiation and oxidizing agents. Traditionally, Escherichia coli and Saccharomyces cerevisiae have been the two platforms of choice for engineering micro-organisms for biotechnological applications, because they are well understood and easy to work with. However, in recent years, researchers have begun using Deinococcus spp in biotechnologies and bioremediation due to their specific ability to grow and express novel engineered functions. More recently, the sequencing of several Deinococcus spp and comparative genomic analysis have provided new insight into the potential of this genus. Features such as the accumulation of genes encoding cell cleaning systems that eliminate organic and inorganic cell toxic components are widespread among Deinococcus spp. Other features such as the ability to degrade and metabolize sugars and polymeric sugars make Deinococcus spp. an attractive alternative for use in industrial biotechnology.

Microencapsulation of probiotic bacteria Lactobacillus plantarum 15HN using alginate-psyllium-fenugreek polymeric blends

Abstract: Aims Investigation on the use of herbal-based biopolymers for probiotic-Lactobacillus plantarum 15HN-encapsulation is presented. The objectives are to enhance its oral delivery, colonic release and survival rate of these probiotic cultures in gastrointestinal environment. Methods and Results Nine types of herbal-based polymers blend with different concentration of alginate alone or mixed with psyllium and fenugreek was used as candidate for encapsulation matrix by applying a simple extrusion method. All the blend formulations recorded high encapsulation efficiency at value >98%. The survival rate of viable probiotic cells under both low pH and high bile salt conditions was also high with value above 80% in 2% (w/v) alginate, alginate+psyllium (1·5 + 0·5%) blend and alginate+fenugreek (1·5 + 0·5%) blend as compared to other polymer formulations and nonencapsulated cells. Their release occurred after 2 h in colonic condition and sustained until the 12th hour incubation period. A value added prebiotic effect was observed in (1·5 + 0·5%) alginate-psyllium formulation. Conclusions The high encapsulation efficiency, high viability of cell in low pH, high bile salt and the sustained release rates of probiotic cells in colonic condition during storage time was also observed for these herbal gel formulations. Significance and Impact of the Study Herbal-based biopolymers offer added advantages of being prebiotic towards the enhancement of probiotic bacterial growth in the gastrointestinal environment.

Production of adipic acid by the native-occurring pathway in Thermobifida fusca B6

Abstract: Aims To identify the native-occurring adipate pathway in a previously engineered Thermobifida fusca B6 strain and optimize the adipic acid production of this strain on glucose and corncob. Methods and results The native-occurring adipate pathway in T. fusca B6 was identified to be the reverse adipate degradation pathway, including five steps: β-ketothiolase (Tfu_0875), 3-hydroxyacyl-CoA dehydrogenase (Tfu_2399), 3-hydroxyadipyl-CoA dehydrogenase (Tfu_0067), 5-Carboxy-2-pentenoyl-CoA reductase (Tfu_1647) and succinyl-CoA synthetase (Tfu_2576, Tfu_2577). The cell lysates of T. fusca wild-type strain with the addition of the Tfu_1647 protein produced trace adipic acid, while no adipic acid was produced in the absence of this protein. The above results prove that the low expression of Tfu_1647 in the wild-type strain was the reason why it did not produce any adipic acid. We then demonstrated that in T. fusca B6, the maximal titre of adipic acid on 50 g l(-1) glucose was 2·23 g l(-1) with 0·045 g g(-1) -glucose yield and 0·22 g l(-1) adipic acid was produced from 19·38 g l(-1) milled corncob. Conclusions The reverse adipate degradation pathway was found to be responsible for the adipate synthesis in T. fusca B6 and Tfu_1647 was the regulatory node on this pathway. Significance and impact of the study Thermobifida fusca B6 was the first reported micro-organism using its native-occurring pathway to accumulate adipic acid. It had the highest reported yield and titre of adipic acid so far.

Survival and persistence of Bacillus clausii in the human gastrointestinal tract following oral administration as spore-based probiotic formulation.

Abstract: AIMS: This study aimed to investigate the fate of Bacillus clausii spores orally administered as lyophilized or liquid formulation to healthy volunteers. METHODS AND RESULTS: The study was a randomized, open‐label, cross‐over trial in which two commercial probiotic formulations containing spores of four antibiotic‐resistant B. clausii strains (OC, NR, SIN, T) were given as a single dose administration. Faecal B. clausii units of each strain were counted on selective media and extrapolated for the total weight of evacuated faeces. RAPD‐PCR typing was used to confirm B. clausii identification. Bacillus clausii was found alive in faeces for up to 12 days. In some volunteers, the recovered amount of OC, NR or SIN was higher than the number of administered spores. Bioequivalence among the two formulations was demonstrated. CONCLUSIONS: Bacillus clausii spores survive transit through the human gastrointestinal tract. They can undergo germination, outgrowth and multiplication as vegetative forms. Bacillus clausii strains can have different ability to survive in the intestinal environment. Bacillus clausii spores administered as liquid suspension or lyophilized form behave similarly in vivo. SIGNIFICANCE AND IMPACT OF THE STUDY: This work contributes towards a better understanding of the behaviour of B. clausii spores as probiotics.

Q fever through consumption of unpasteurised milk and milk products – a risk profile and exposure assessment

Abstract: Q fever is a zoonotic disease caused by the bacterium Coxiella burnetii which is endemic in cattle, sheep and goats in much of the world, including the United Kingdom (UK). There is some epidemiological evidence that a small proportion of cases in the developed world may arise from consumption of unpasteurised milk with less evidence for milk products such as cheese. Long maturation at low pH may give some inactivation in hard cheese, and viable C. burnetii are rarely detected in unpasteurised cheese compared to unpasteurised milk. Simulations presented here predict that the probability of exposure per person to one or more C. burnetii through the daily cumulative consumption of raw milk in the UK is 04203. For those positive exposures, the average level of exposure predicted is high at 1266 guinea pig intraperitoneal infectious dose 50% units (GP_IP_ID50) per person per day. However, in the absence of human dose–response data, the case is made that the GP_IP_ID50 unit represents a very low risk through the oral route. The available evidence suggests that the risks from C. burnetii through consumption of unpasteurised milk and milk products (including cheese) are not negligible but they are lower in comparison to transmission via inhalation of aerosols from parturient products and livestock contact.

Evaluation of phosphate‐solubilizing bacteria on the growth and grain yield of rice (Oryza sativa L.) cropped in northern Iran

Abstract: Aims This study aimed to evaluate the efficiency of four phosphate-solubilizing bacteria (PSB) on the growth and yield of rice under different soil conditions. Methods Bacterial strains were Rahnella aquatillis (KM977991), Enterobacter sp. (KM977992), Pseudomonas fluorescens and Pseudomonas putida. These studies were conducted on different rice cultivars (‘Shiroodi’, ‘Tarom’ and ‘Tarom Hashemi’) in both pot and field experiments. Measurements started from transplanting and continued throughout the growing season in field experiments. Results Single PSB inoculations in field trials increased grain yield, biological yield, total number of stems hill−1, number of panicles hill−1 and plant height by 8·50–26·9%, 12·4–30·9%, 20·3–38·7%, 22·1–36·1% and 0·85–3·35% in experiment 1, by 7·74–14·7%, 4·22–12·6%, 6·67–16·7%, 4·0–15·4% and 3·15–4·20% in experiment 2 and by 23·4–37%, 16·1–36·4%, 30·2–39·1%, 28·8–34% and 2·11–4·55% in experiment 3, respectively, compared to the control. Our results indicate that the application of triple super phosphate together with PSB inoculations resulted in reducing the use of chemical fertilizers (about 67%) and increasing fertilizer use efficiency. Conclusions This study clearly indicates that these PSBs can be used as biofertilizers in ecological rice agricultural systems. Significance and Impact of the Study To the best of our knowledge, this is first report on the association of Rahnella aquatilis with rice and also the application of a mathematical model to evaluate the effect of PSBs on rice growth.

Metabolic engineering for isoprenoid-based biofuel production

Abstract: Summary Sustainable economic and industrial growth is the need of the hour and it requires renewable energy resources having better performance and compatibility with existing fuel infrastructure from biological routes. Isoprenoids (C ≥ 5) can be a potential alternative due to their diverse nature and physiochemical properties similar to that of petroleum based fuels. In the past decade, extensive research has been done to utilize metabolic engineering strategies in micro-organisms primarily, (i) to overcome the limitations associated with their natural and non-natural production and (ii) to develop commercially competent microbial strain for isoprenoid-based biofuel production. This review briefly describes the engineered isoprenoid biosynthetic pathways in well-characterized microbial systems for the production of several isoprenoid-based biofuels and fuel precursors.

Effects of graded levels of tannin-containing tropical tree leaves on in vitro rumen fermentation, total protozoa and methane production.

Abstract: Aims This study was carried out to determine the effect of graded levels of tannin-containing tropical tree leaves, Autocarpus integrifolis, Azardirachta indica and Ficus bengalensis, on the in vitro rumen fermentation pattern, total protozoa and methane suppression in order to establish the optimum dose of these leaves for inclusion in the ruminant diets. Methods and Results The air-dried and ground samples of Au. integrifolis, Az. indica and Ficus bengalensis were subjected to in vitro incubation using 30 ml buffered rumen fluid at 0, 2·5, 5·0, 10·0, 15·0, 20·0, 25·0 and 30·0% (dry matter refers to moisture-free basis) of a total mixed ration (TMR: refers to mixture of roughage and concentrate containing cereals and oil cakes) devoid of tannin. The TMR for the experimental incubation was prepared by mixing 40 parts of ground Elusine coracana straw as roughage source with 60 parts of concentrate mixture. The leaves contained an average 130 g kg−1 CP with 7·0 MJ of ME kg−1 DM. The average neutral detergent fibre (NDF) content was 5·0%) reduced TVFA concentration. Protozoa (cells per mL) were similar at all levels of inclusion with Au. integrifolis, but reduced in case of F. bengalensis and Az. indica. As the level of tannin increased in the incubation medium, there was a linear reduction in methane concentration. Highest methane reduction (%) was recorded in incubations supplemented with Az. indica (61·5) followed by F. bengalensis (46·8) and Au. integrifolis (30·3). Conclusions It was established from this study that tropical leaves of F. bengalensis, Au. integrifolis and Az. indica suppress methanogenesis. Significance and Impact of the Study Ficus bengalensis, Au. integrifolis and Az. indica leaves are of interest in the enteric methane ameliorative strategies. Total mixed ration containing 10–15% ground F. bengalensis or Au. integrifolis or Az. indica leaves could be fed to ruminants to reduce enteric methanogenesis. However, in vivo trials need to be carried out to validate these in vitro results.