Complete and Draft Genome Sequences of 52 Bacillus and Priestia Strains Isolated from West African Fermentations and 26 Reference Strains from a Public Culture Collection.

The whole genomes of 78 Bacillus and Priestia strains isolated from West African fermented foods (n = 52) or acquired from a public culture collection (n = 26) were sequenced using long-read sequencing and assembled into draft (n = 32) and complete (n = 46) genomes, allowing comparative genomics and taxonomic assignment of these strains with putative uses in fermented foods.

Machine learning methods for the detection of explosives, drugs and precursor chemicals gathered using a colorimetric sniffer sensor.

Colorimetric sensing technology for the detection of explosives, drugs, and their precursor chemicals is an important and effective approach. In this work, we use various machine learning models to detect these substances from colorimetric sensing experiments conducted in controlled environments. The detection experiments based on the response of a colorimetric chip containing 26 chemo-responsive dyes indicate that homemade explosives (HMEs) such as hexamethylene triperoxide diamine (HMTD), triacetone triperoxide (TATP), and methyl ethyl ketone peroxide (MEKP) used in improvised explosives devices are detected with true positive rate (TPR) of 70-75%, 73-90% and 60-82% respectively. Time series classifiers such as Convolutional Neural Networks (CNN) are explored, and the results indicate that improvements can be achieved with the use of kinetics of the chemical responses. The use of CNNs is limited, however, to scenarios where a large number of measurements, typically in the range of a few hundred, of each analyte are available. Feature selection of important dyes using the Group Lasso (GPLASSO) algorithm indicated that certain dyes are more important in discrimination of an analyte from ambient air. This information could be used for optimizing the colorimetric sensor and extend the detection to more analytes.

Transport of citrate and polymer coated gold nanoparticles (AuNPs) in porous media: Effect of surface property and Darcy velocity.

With the release of nanoparticles (NPs) into the subsurface, it is imperative to better understand the fate and transport of NPs in porous media. Three types of stable AuNPs were used as model NPs to investigate the impact of surface coatings (type and coverage) and water velocity on the NP transport in a porous media (column studies). The NPs were electrostatic stabilized citrate AuNPs and sterically stabilized AuNPs with amphiphilic block co-polymer (PVA-COOH) in two particle/polymer ratios (weak vs. strong stabilization). The citrate AuNPs transport was sensitive to ionic changes in the mixing front of the plume, where destabilization occurred, and will therefore depend on the size/type of release. Blocking of deposition sites by aggregates was seen to facilitate transport, whereby a higher flow velocity (larger shadow zone) also resulted in better transport. The polymeric surface coating had great impact with steric repulsion as a main force contributing to the transport of NPs in the porous media. Sufficient polymer coating was crucial to obtain highly unfavorable attachment conditions (very low α) where the enhanced NP mobility was independent of the water velocity (comparable to solute tracer). Without sufficient steric stabilization, the transport and recovery was significantly reduced compared to the solute tracer, but increased with increasing water velocity. This highlights the importance of sufficient surface coating to achieve enhanced mobility, but also the increased risk of spreading to down-gradient receptors. For the (weakly) sterically stabilized NPs, the loss of polymer through ligand exchange with the porous media negates transport.

Nanoimprinting and tapering of chalcogenide photonic crystal fibers for cascaded supercontinuum generation.

Improved long-wavelength transmission and supercontinuum (SC) generation is demonstrated by antireflective (AR) nanoimprinting and tapering of chalcogenide photonic crystal fibers (PCFs). Using a SC source input spanning from 1 to 4.2 µm, the total transmission of a 15 µm core diameter PCF was improved from ∼53% to ∼74% by nanoimprinting of AR structures on both input and output facets of the fiber. Through a combined effect of reduced reflection and redshifting of the spectrum to 5 µm, the relative transmission of light >3.5 µm in the same fiber was increased by 60.2%. Further extension of the spectrum to 8 µm was achieved using tapered fibers. The spectral broadening dynamics and output power were investigated using different taper parameters and pulse repetition rates.

Nanoimprinting reflow modified moth-eye structures in chalcogenide glass for enhanced broadband antireflection in the mid-infrared.

We report on the progress towards developing a new method for fabricating more efficient, broadband antireflective (AR) moth-eye structures in As2Se3 via a direct nanoimprinting technique. Thermal reflow is used during mold fabrication to reshape a conventional deep-ultraviolet lithography in order to promote a pattern transfer of "secant ogive"-like moth-eye structures. Once replicated, structures modified by reflow displayed greater AR efficiency compared to structures replicated by a conventional mold, achieving the highest spectrum-averaged transmittance improvement of 12.36% from 3.3 to 12 µm.

Mobility of electrostatically and sterically stabilized gold nanoparticles (AuNPs) in saturated porous media.

The stability of gold nanoparticles (AuNPs) stabilized electrostatically with citrate or (electro)sterically by commercially available amphiphilic block copolymers (PVP-VA or PVA-COOH) was studied under various physicochemical conditions. Subsequently, the mobility of the AuNPs in porous media (sand) was investigated in column studies under environmental relevant physicochemical conditions. Electrostatically stabilized AuNPs were unstable under most physicochemical conditions due to the compression of the electrical double layer. Consequently, aggregation and deposition rapidly immobilized the AuNPs. Sterically stabilized AuNPs showed significantly less sensitivity towards changes in the physicochemical conditions with high stability, high mobility with negligible retardation, and particle deposition rate coefficients ranging an order of magnitude (1.5 × 10-3 to 1.5 × 10-2 min-1) depending on the type and amount of stabilizer, and thereby the surface coverage and attachment affinity. The transport of sterically stabilized AuNPs is facilitated by reversible deposition in shallow energy minima with continuous reentrainment and blocking of available attachment sites by deposited AuNPs. The stability and mobility of NPs in the environment will thereby be highly dependent on the specific stabilizing agent and variations in the coverage on the NP. Under the given experimental conditions, transport distances of the most mobile AuNPs of up to 20 m is expected. Due to their size-specific plasmonic properties, the easily detectable AuNPs are proposed as potential model or tracer particles for studying transport of various stabilized NPs under environmental conditions.

Prevalence of foodborne pathogens in food from selected African countries - A meta-analysis.

Food safety information in the African region is insufficient and fragmented due to lack of surveillance, documentation and reporting, thereby resulting in inefficient utilization of resources, duplication of activities, and lack of synergy among the countries of the region. This paper reviews the prevalence of foodborne pathogens in seven African countries (Benin, Botswana, Ghana, Kenya, Nigeria, Sudan and Uganda) from papers in regional or international journals published between January 2000 and December 2015. One hundred and sixteen publications that dealt with food microbiology were reviewed for general analysis, while 66 papers on contamination of pathogenic bacteria were used for meta-analysis of prevalence. The food items were split into two categories: raw foods and ready-to-eat (RTE) foods (including street food and beverages) for meta-analysis. Majority of the reviewed studies (67.2%, 78/116) dealt with food of animal origin: 38.8% for meat and eggs, 17.2% for dairy products and 11.2% for aquatic products. Only 8.6% examined foods of plant origin (fruits and vegetables). The remaining 24.1% was the composite RTE food and beverages. Enterobacteriaceae, Escherichia coli, Salmonella, Staphylococcus aureus and Listeria monocytogenes were the most frequently reported organisms in those studies. Although the data were highly heterogeneous, a striking feature is high prevalence of the major pathogens in RTE foods, almost as high as in raw foods. E. coli averaged at 37.6% in raw foods and 31.6% in RTE foods. The corresponding prevalence for Salmonella was 19.9% vs 21.7%; S. aureus, 27.8% vs 25.1% and L. monocytogenes, 19.5% vs 6.7%. The average prevalence of foodborne pathogens in these countries was 34.2% (29.0-39.3%). Differences in food types as well as non-uniform protocols for sampling and identification might have contributed to high heterogeneity (I2 >97%) although some high prevalence data could be factual with extensive varieties of raw and RTE foods. Need for improved hygienic practices in handling of raw or RTE foods are suggested. Implementation of surveillance programs that use uniform laboratory protocols across the region could give homogeneous results.

Selective removal of heavy metal ions by disulfide linked polymer networks.

Heavy metal contaminated surface water is one of the oldest pollution problems, which is critical to ecosystems and human health. We devised disulfide linked polymer networks and employed as a sorbent for removing heavy metal ions from contaminated water. Although the polymer network material has a moderate surface area, it demonstrated cadmium removal efficiency equivalent to highly porous activated carbon while it showed 16 times faster sorption kinetics compared to activated carbon, owing to the high affinity of cadmium towards disulfide and thiol functionality in the polymer network. The metal sorption mechanism on polymer network was studied by sorption kinetics, effect of pH, and metal complexation. We observed that the metal ions-copper, cadmium, and zinc showed high binding affinity in polymer network, even in the presence of competing cations like calcium in water.

In situ SU-8 silver nanocomposites.

Nanocomposite materials containing metal nanoparticles are of considerable interest in photonics and optoelectronics applications. However, device fabrication of such materials always encounters the challenge of incorporation of preformed nanoparticles into photoresist materials. As a solution to this problem, an easy new method of fabricating silver nanocomposites by an in situ reduction of precursors within the epoxy-based photoresist SU-8 has been developed. AgNO3 dissolved in acetonitrile and mixed with the epoxy-based photoresist SU-8 forms silver nanoparticles primarily during the pre- and post-exposure soft bake steps at 95 °C. A further high-temperature treatment at 300 °C resulted in the formation of densely homogeneously distributed silver nanoparticles in the photoresist matrix. No particle growth or agglomeration of nanoparticles is observed at this point. The reported new in situ silver nanocomposite materials can be spin coated as homogeneous thin films and structured by using UV lithography. A resolution of 5 µm is achieved in the lithographic process. The UV exposure time is found to be independent of the nanoparticle concentration. The fabricated silver nanocomposites exhibit high plasmonic responses suitable for the development of new optoelectronic and optical sensing devices.

Micro-flow-injection analysis (µFIA) immunoassay of herbicide residue 2,6-dichlorobenzamide ­ towards automated at-line monitoring using modular microfluidics.

As a part of developing new systems for continuously monitoring the presence of pesticides in groundwater, a microfluidic amperometric immunosensor was developed for detecting the herbicide residue 2,6-dichlorobenzamide (BAM) in water. A competitive immunosorbent assay served as the sensing mechanism and amperometry was applied for detection. Both the immunoreaction chip (IRC) and detection (D) unit are integrated on a modular microfluidic platform with in-built micro-flow-injection analysis (µFIA) function. The immunosorbent, immobilized in the channel of the IRC, was found to have high long-term stability and withstand many regeneration cycles, both of which are key requirements for systems utilized in continuous monitoring. The IRC was regenerated during 51 cycles in a heterogeneous competitive assay out of which 27 were without the analyte (the highest possible signal level) in order to assess the regeneration capability of the immunosorbent. Detection of BAM standard solutions was performed in the concentration range from 62.5 µg L(-1) to 0.0008 µg L(-1). Non-linear regression of the data using the four-parameter logistic equation generated a sigmoidal standard curve showing an IC50 value (concentration that reduces the signal by 50%) of 0.25 µg L(-1). The strongest signal variation is observed in the concentration range between 0.02 and 2.5 µg L(-1), which includes the 0.1 µg L(-1) threshold limit set by the European Commission for BAM in drinking water. The presented results demonstrate the potential of the constructed µFIA immunosensor as an at-line monitoring system for controlling the quality of ground water supply.

Assessment of interplay between UV wavelengths, material surfaces and food residues in open surface hygiene validation.

The use of UV-visible radiation for detecting invisible residue on different surfaces as a means of validating cleanliness was investigated. Wavelengths at 365, 395, 435, 445, 470 and 490 nm from a monochromator were used to detect residues of beef, chicken, apple, mango and skim milk. These were on three surfaces: aluminium, fibre re-enforced plastic (FRP; Q-Liner®) and stainless steel, pre- and post a cleaning step using commercial detergent. The area covered by residues as detected by specific wavelengths was compared statistically. The sensitivity of the wavelengths for detection differed significantly (p < 0.05) for various residues depending on the material surfaces. Generally, wavelengths 365-445 nm were consistently able to illuminate all residue before cleaning, though sensitivity varied, while 490 nm showed more of the surface structural features instead of residue. The 365-395 nm wavelengths were significantly more sensitive (p < 0.05) for detecting beef and chicken residues on aluminium and stainless steel both before and after cleaning. The 435-445 nm wavelengths were significantly more sensitive for detecting apple and mango residues on the FRP both before and after cleaning. It is important when UV-systems are used as real-time tools for assessing cleanliness of surfaces that the surface materials being illuminated are taken into account in the choice of lamp wavelength, in addition to expected residue. This will ensure higher confidence in results during the use of UV-light for real-time hygiene validation of surfaces.

Synbiotic Lactobacillus acidophilus NCFM and cellobiose does not affect human gut bacterial diversity but increases abundance of lactobacilli, bifidobacteria and branched-chain fatty acids: a randomized, double-blinded cross-over trial.

Probiotics, prebiotics, and combinations thereof, that is synbiotics, have been reported to modulate gut microbiota of humans. In this study, effects of a novel synbiotic on the composition and metabolic activity of human gut microbiota were investigated. Healthy volunteers (n = 18) were enrolled in a double-blinded, randomized, and placebo-controlled cross-over study and received synbiotic [Lactobacillus acidophilus NCFM (10(9) CFU) and cellobiose (5 g)] or placebo daily for 3 weeks. Fecal samples were collected and lactobacilli numbers were quantified by qPCR. Furthermore, 454 tag-encoded amplicon pyrosequencing was used to monitor the effect of synbiotic on the composition of the microbiota. The synbiotic increased levels of Lactobacillus spp. and relative abundances of the genera Bifidobacterium, Collinsella, and Eubacterium while the genus Dialister was decreased (P < 0.05). No other effects were found on microbiota composition. Remarkably, however, the synbiotic increased concentrations of branched-chain fatty acids, measured by gas chromatography, while short-chain fatty acids were not affected.

Determination of yeast diversity in ogi, mawè, gowé and tchoukoutou by using culture-dependent and -independent methods.

The maize based ogi and mawè and the sorghum based gowé and tchoukoutou are traditional, spontaneously fermented products widely consumed by the population of Benin (West Africa). Yeast occurrence in the products, as sold on local markets at different locations, was studied using a combination of culture-dependent and independent methods. Number of yeasts is varied from 3.75 log10 colony forming units (cfu)/g for ogi to 5.60 log10 cfu/g for tchoukoutou. Isolated yeasts (236) were identified based on different migration profiles on denaturing gradient gel electrophoresis (DGGE) and 26S rRNA gene sequencing. Candida krusei was the yeast most frequently isolated with strongest predominance in the maize based products. Other predominant yeast present at equal or lower incidence were Clavispora lusitaniae and Saccharomyces cerevisiae in ogi and mawè, Cl. lusitaniae, Candida tropicalis and Kluyveromyces marxianus in gowè and Cl. lusitaniae, S. cerevisiae and Candida rugosa in tchoukoutou. Grouping of C. krusei isolates (164) by rep-PCR analysis indicated that several biotypes were involved in fermentation of the four products. The DGGE analysis on the DNA directly extracted from the food matrices demonstrated the presence of Dekkera bruxellensis and Debaryomyces hansenii, not detected by the culture-based approach. This is the first study combining culture-dependent and independent methods to reveal predominant yeast species and biotypes in traditional foods from Benin.

Yeast dynamics during spontaneous fermentation of mawè and tchoukoutou, two traditional products from Benin.

Mawè and tchoukoutou are two traditional fermented foods largely consumed in Benin, West Africa. Their preparations remain as a house art and they are the result of spontaneous fermentation processes. In this study, dynamics of the yeast populations occurring during spontaneous fermentations of mawè and tchoukoutou were investigated using both culture-dependent and -independent approaches. For each product, two productions were followed. Samples were taken at different fermentation times and yeasts were isolated, resulting in the collection of 177 isolates. They were identified by the PCR-DGGE technique followed by the sequencing of the D1/D2 domain of the 26S rRNA gene. The predominant yeast species identified were typed by rep-PCR. Candida krusei was the predominant yeast species in mawè fermentation followed by Candida glabrata and Kluyveromyces marxianus. Other yeast species were detected in lower numbers. The yeast successions that took place during mawè fermentation lead to a final population comprising Saccharomyces cerevisiae, C. krusei and K. marxianus. The yeast populations dominating the fermentation of tchoukoutou were found to consist of S. cerevisiae, almost exclusively. Other yeast species were detected in the early stages of fermentation. For the predominant species a succession of biotypes was demonstrated by rep-PCR for the fermentation of both products. The direct analysis at DNA and RNA levels in the case of mawè did not reveal any other species except those already identified by culture-based analysis. On the other hand, for tchoukoutou, four species were identified that were not detected by the culture-based approach. The spontaneous fermentation of mawè and tchoukoutou in the end were dominated by a few autochthonous species.

Biodiversity of aerobic endospore-forming bacterial species occurring in Yanyanku and Ikpiru, fermented seeds of Hibiscus sabdariffa used to produce food condiments in Benin.

Yanyanku and Ikpiru made by the fermentation of Malcavene bean (Hibiscus sabdariffa) are used as functional additives for Parkia biglobosa seed fermentations in Benin. A total of 355 aerobic endospore-forming bacteria (AEFB) isolated from Yanyanku and Ikpiru produced in northern and southern Benin were identified using phenotypic and genotypic methods, including GTG5-PCR, M13-PCR, 16S rRNA, gyrA and gyrB gene sequencing. Generally, the same 5-6 species of the genus Bacillus predominated: Bacillus subtilis (17-41% of isolates), Bacillus cereus (8-39%), Bacillus amyloliquefaciens (9-22%), Bacillus licheniformis (3-26%), Bacillus safensis (8-19%) and Bacillus altitudinis (0-19%). Bacillus aryabhattai, Bacillus flexus, and Bacillus circulans (0-2%), and species of the genera Lysinibacillus (0-14%), Paenibacillus (0-13%), Brevibacillus (0-4%), and Aneurinibacillus (0-3%) occurred sporadically. The diarrheal toxin encoding genes cytK-1, cytK-2, hblA, hblC, and hblD were present in 0%, 91% 15%, 34% and 35% of B. cereus isolates, respectively. 9% of them harbored the emetic toxin genetic determinant, cesB. This study is the first to identify the AEFB of Yanyanku and Ikpiru to species level and perform a safety evaluation based on toxin gene detections. We further suggest, that the gyrA gene can be used for differentiating the closely related species Bacillus pumilus and B. safensis.

Co-production of surfactin and a novel bacteriocin by Bacillus subtilis subsp. subtilis H4 isolated from Bikalga, an African alkaline Hibiscus sabdariffa seed fermented condiment.

Bikalga is a Hibiscus sabdariffa seed fermented condiment widely consumed in Burkina Faso and neighboring countries. The fermentation is dominated by Bacillus subtilis group species. Ten B. subtilis subsp. subtilis (six isolates) and Bacillus licheniformis (four isolates) isolated from traditional Bikalga were examined for their antimicrobial activity against a panel of 36 indicator organisms including Gram-positive and Gram-negative bacteria and yeasts. The Bacillus spp. isolates showed variable inhibitory abilities depending on the method used. Both Gram-positive and Gram-negative bacteria were inhibited in the agar spot assay while only Gram-positive pathogens were inhibited in the agar well diffusion assay. Cell free supernatants (CFS) of pure cultures of 3 B. subtilis subsp. subtilis (G2, H4 and F1) strains inhibited growth of Listeria monocytogenes, Micrococcus luteus, Staphylococcus aureus and Bacillus cereus, while CFS of 2 B. licheniformis (E3 and F9) strains only inhibited M. luteus. The antimicrobial substance(s) produced by B. subtilis subsp. subtilis H4 was further characterized. The antimicrobial substance(s) produced by H4 was detected from mid-exponential growth phase. The activity was sensitive to protease and trypsin, but resistant to the proteolytic action of proteinase K and papain. Treatment with α-amylase and lipase II resulted in a complete loss of antimicrobial effect, indicating that a sugar moiety and lipid moiety are necessary for the activity. Treatment with mercapto-ethanol resulted in a significant loss, indicative of the presence of disulfide bridges. The antimicrobial activity of H4 was heat resistant and active at pH3-10. PCR detection of yiwB, sboA, spoX, albA and spaS, etnS genes and genes coding for surfactins and plipastatins (fengycins) indicated a potential for subtilosin, subtilin and lipopeptide production, respectively. Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) was carried out and a single band of approximately 4kDa had antimicrobial activity. Ultra high performance liquid chromatography-time of flight mass spectrometry (UHPLC-TOFMS) analysis of the 4kDa band allowed identification of surfactin and a protein with a monoisotopic mass of 3346.59Da, which is dissimilar in size to subtilosin and subtilin. Surfactin is a cyclic lipoheptapeptide, which contains a ß-hydroxy fatty acid, but no di-sulfide bridges or sugar residues. The complete loss of activity upon amylase treatment indicates that surfactin was not responsible for the observed antimicrobial effect. However, it cannot completely be ruled out that surfactin acts synergistically with the detected protein, though further investigations are needed to confirm this.

Effect of Lactobacillus salivarius Ls-33 on fecal microbiota in obese adolescents.

BACKGROUND & AIMS: This study is a part of the clinical trials with probiotic bacterium Lactobacillus salivarius Ls-33 conducted in obese adolescents. Previously reported clinical studies showed no effect of Ls-33 consumption on the metabolic syndrome in the subject group. The aim of the study was to investigate the impact of L. salivarius Ls-33 on fecal microbiota in obese adolescents. METHODS: The study was a double-blinded intervention with 50 subjects randomized to intake of L. salivarius Ls-33 or placebo for 12 weeks. The fecal microbiota was assessed by real-time quantitative PCR before and after intervention. Concentrations of fecal short chain fatty acids were determined using gas chromatography. RESULTS: Ratios of Bacteroides-Prevotella-Porphyromonas group to Firmicutes belonging bacteria, including Clostridium cluster XIV, Blautia coccoides_Eubacteria rectale group and Roseburia intestinalis, were significantly increased (p ≤ 0.05) after administration of Ls-33. The cell numbers of fecal bacteria, including the groups above as well as Clostridium cluster I, Clostridium cluster IV, Faecalibacterium prausnitzii, Enterobacteriaceae, Enterococcus, the Lactobacillus group and Bifidobacterium were not significantly altered by intervention. Similarly, short chain fatty acids remained unaffected. CONCLUSION: L. salivarius Ls-33 might modify the fecal microbiota in obese adolescents in a way not related to metabolic syndrome. CLINICAL TRIAL NUMBER: NCT 01020617.

Inhibition of Bacillus cereus growth by bacteriocin producing Bacillus subtilis isolated from fermented baobab seeds (maari) is substrate dependent.

Maari is a spontaneously alkaline fermented food condiment made from baobab tree seeds. Due to the spontaneous nature of maari fermentations growth of the opportunistic human pathogen Bacillus cereus is occasionally observed. Bacillus subtilis strains are important for alkaline seed fermentations because of their enzymatic activities contributing to desirable texture, flavor and pH development. Some B. subtilis strains have antimicrobial properties against B. cereus. In the present work, three bacteriocin producing B. subtilis strains (B3, B122 and B222) isolated from maari were tested. The production of antimicrobial activity by the three strains was found to be greatly influenced by the substrate. All three B. subtilis strains produced antimicrobial activity against B. cereus NVH391-98 in BHI broth as determined by the agar well diffusion assay, whereas no antimicrobial activity was detected in whole cooked baobab seeds and in 10% (w/v) grinded baobab seeds. Incorporation of BHI with up to 5% (w/w) grinded baobab seeds enhanced the antimicrobial activity of B. subtilis compared with pure BHI in a strain dependent manner. Incorporation of BHI with 50% (w/w) baobab grinded seeds decreased the antimicrobial activity. Addition of the inorganic salts FeCl3, MgSO4 and MnSO4 has previously been reported to increase bacteriocin production of B. subtilis, but the addition of these salts to 10% (w/v) grinded baobab seed broth did not cause antimicrobial activity. Survival of B. cereus NVH391-98 in co-culture with B. subtilis was tested in BHI broth, 10% (w/v) grinded baobab seed based broth and during baobab seed fermentation to produce maari. B. cereus NVH391-98 grew well in all three substrates in mono-culture. All the 3 B. subtilis strains were able to decrease B. cereus NVH391-98 to levels below the detection limit (<10 CFU/ml) in BHI, but not in baobab seed based substrates, even though the outgrowth of B. cereus NVH391-98 was delayed by up to 40 h. In conclusion, production of antimicrobial activity by the investigated B. subtilis strains is highly substrate-specific and strain-specific. The three B. subtilis strains delayed but did not prevent B. cereus outgrowth during baobab seed fermentations. Maari is produced through mixed microbial population fermentations. B. subtilis co-starter culture candidates originating from maari which are able to prevent pathogen outgrowth remain to be identified.

Screening for Bacillus subtilis group isolates that degrade cyanogens at pH 4.5-5.0.

Cyanogenic food crops abound in nature with important crops like cassava forming the staple food for over half a billion people. Detoxification by hydrolysis of cassava cyanogenic glycosides often involves acid fermentation, and in some of these processes Bacillus species are encountered. Forty Bacillus spp. (20 Bacillus subtilis, 11 Bacillus licheniformis, 7 Bacillus sonorensis, 2 Bacillus cereus) isolated from acid fermented primary starters to produce Gergoush, a Sudanese fermented snack, were screened for their ability to grow and to hydrolyze linamarin, the major cyanogen found in cassava at pH levels below 5.0; also the cyanogen amygdalin was assessed. The B. subtilis isolates grew in both HCl and lactic acid environments from pH 4.5-6.0 while being able to break down the cyanogenic glycosides. The B. licheniformis and B. sonorensis isolates grew and degraded cyanogens at pH 5.0 in a HCl environment, while two B. cereus isolates used in the study showed no breakdown reaction under all conditions tested. One B. subtilis isolate was observed to have substrate specificity between the breakdown of linamarin and amygdalin. We conclude that some Bacillus spp. isolates are important in the microbiological breakdown of cyanogens in cassava fermentations even at pH 4.5-5.0 though further investigations are required.

The effect of selected synbiotics on microbial composition and short-chain fatty acid production in a model system of the human colon.

BACKGROUND: Prebiotics, probiotics and synbiotics can be used to modulate both the composition and activity of the gut microbiota and thereby potentially affecting host health beneficially. The aim of this study was to investigate the effects of eight synbiotic combinations on the composition and activity of human fecal microbiota using a four-stage semicontinuous model system of the human colon. METHODS AND FINDINGS: Carbohydrates were selected by their ability to enhance growth of the probiotic bacteria Lactobacillus acidophilus NCFM (NCFM) and Bifidobacterium animalis subsp. lactis Bl-04 (Bl-04) under laboratory conditions. The most effective carbohydrates for each probiotic were further investigated, using the colonic model, for the ability to support growth of the probiotic bacteria, influence the composition of the microbiota and stimulate formation of short-chain fatty acids (SCFA).The following combinations were studied: NCFM with isomaltulose, cellobiose, raffinose and an oat ß-glucan hydrolysate (OBGH) and Bl-04 with melibiose, xylobiose, raffinose and maltotriose. All carbohydrates showed capable of increasing levels of NCFM and Bl-04 during fermentations in the colonic model by 10(3)-10(4) fold and 10-10(2) fold, respectively. Also the synbiotic combinations decreased the modified ratio of Bacteroidetes/Firmicutes (calculated using qPCR results for Bacteroides-Prevotella-Porphyromonas group, Clostridium perfringens cluster I, Clostridium coccoides - Eubacterium rectale group and Clostridial cluster XIV) as well as significantly increasing SCFA levels, especially acetic and butyric acid, by three to eight fold, as compared to the controls. The decreases in the modified ratio of Bacteroidetes/Firmicutes were found to be correlated to increases in acetic and butyric acid (p=0.04 and p=0.03, respectively). CONCLUSIONS: The results of this study show that all synbiotic combinations investigated are able to shift the predominant bacteria and the production of SCFA of fecal microbiota in a model system of the human colon, thereby potentially being able to manipulate the microbiota in a way connected to human health.