Formulation and stabilization of an Arthrobacter strain with good storage stability and 4-chlorophenol-degradation activity for bioremediation.

Chlorophenols are widespread and of environmental concern due to their toxic and carcinogenic properties. Development of less costly and less technically challenging remediation methods are needed; therefore, we developed a formulation based on micronized vermiculite that, when air-dried, resulted in a granular product containing the 4-chlorophenol (4-CP)-degrading Gram-positive bacterium Arthrobacter chlorophenolicus A6. This formulation and stabilization method yielded survival rates of about 60% that remained stable in storage for at least 3 months at 4 °C. The 4-CP degradation by the formulated and desiccated A. chlorophenolicus A6 cells was compared to that of freshly grown cells in controlled-environment soil microcosms. The stabilized cells degraded 4-CP equally efficient as freshly grown cells in two different set-ups using both hygienized and non-treated soils. The desiccated microbial product was successfully employed in an outdoor pot trial showing its effectiveness under more realistic environmental conditions. No significant phytoremediation effects on 4-CP degradation were observed in the outdoor pot experiment. The 4-CP degradation kinetics from both the microcosms and the outdoor pot trial were used to generate a predictive model of 4-CP biodegradation potentially useful for larger-scale operations, enabling better bioremediation set-ups and saving of resources. This study also opens up the possibility of formulating and stabilizing also other Arthrobacter strains possessing different desirable pollutant-degrading capabilities.

Knockdown of MSI1 inhibits the proliferation of human oral squamous cell carcinoma by inactivating STAT3 signaling.

Musashi RNA­binding protein 1 (MSI1) is highly expressed in several types of cancer; however, its role in oral squamous cell carcinoma (OSCC) remains unknown. The purpose of this study was to investigate the probable mechanism underlying the involvement of MSI1 in OSCC. The results demonstrated that MSI1 was upregulated in OSCC tissues, but not in adjacent healthy tissues. MSI1 silencing resulted in decreased cell proliferative, invasive and migrative capacity. In addition, MSI1 silencing led to cell cycle arrest at the S phase, downregulation of c­Myc and cyclin D1, and upregulation of p21 and p27 levels. Additional studies demonstrated that MSI1 suppression inhibited the activation of signal transducer and activator of transcription 3 (STAT3) signaling. Accordingly, the findings of the present study suggested that MSI1 silencing can suppress OSCC cell proliferation and progression, in part by inhibiting the activation of the c­Myc/STAT3 pathway.

Production of volatile compounds by Rhizopus oligosporus during soybean and barley tempeh fermentation.

Rhizopus oligosporus Saito can ferment soybeans or cereal grains to tempeh, a sliceable cake with improved nutritional properties. Volatiles produced by different R. oligosporus strains grown on malt extract agar (MEA), barley and soybean were investigated. The effect of co-cultivation with Lactobacillus plantarum on the production of volatiles was also studied. Volatile compounds were collected in situ by headspace diffusion and identified by GC-MS. The ten R. oligosporus strains that had different colony morphologies on MEA produced very similar volatile profiles, except for slight variations among the minor volatile compounds (e.g. sesquiterpenes). Likewise, practically no differences in volatile profiles were observed between three of the strains grown on soybeans. In contrast, the R. oligosporus volatile profile on soybean was different from that on barley from the same strain. Co-cultivation with L. plantarum did not influence volatile production by R. oligosporus. The dominant compounds produced on all three substrates were ethanol, acetone, ethyl acetate, 2-butanone, 2-methyl-1-propanol, 3-methyl-1-butanol and 2-methyl-1-butanol. Acetaldehyde and 2-methyl-propanal were also produced on MEA and barley, while 2-pentanone, methyl acetate, 2-butanol and 3-methyl-3-buten-1-ol were observed on soybeans. Ethanol, 2-methyl-1-butanol and 3-methyl-1-butanol were the most abundant volatile compounds produced on MEA and barley, while 2-butanone was the dominant volatile metabolite on soybean. The mushroom odour compounds, 3-octanone and 1-octen-3-ol, were only detected from soybean and soybean tempeh.

Growth of lactic acid bacteria and Rhizopus oligosporus during barley tempeh fermentation.

The zygomycete Rhizopus oligosporus is traditionally used to ferment soybean tempeh, but it is also possible to ferment other legumes and cereals to tempeh. The traditional soybean tempeh harbours a multitude of microorganisms with potentially beneficial or detrimental effects on quality. Lactic acid bacteria (LAB) have positive effects on the safety of soybean tempeh, but the effects of LAB on R. oligosporus growth have not been investigated. We have developed a cereal grain tempeh by fermenting pearled barley with R. oligosporus ATCC 64063. Four LAB species, Lactobacillus plantarum, Lactobacillus fermentum, Lactobacillus reuteri and Lactococcus lactis were assessed for their growth abilities and their effects on R. oligosporus growth during barley tempeh fermentation. Growth of LAB was assayed as colony forming units (cfu), while growth of R. oligosporus was measured as ergosterol content and hyphal length. The two fungal measurements highly correlated (r=0.83, P<0.001, n=90). The ergosterol content of fungal mycelia ranged from 11.7 to 30.1 mg/g fungal dry matter. L. plantarum multiplied from 4.8 to 7.4 log cfu/g dry tempeh and L. fermentum increased from 4.4 to 6.8 log cfu/g during 24 h incubation at 35 degrees C. L. reuteri and L. lactis had significantly slower growth, with increases from 4.8 to 5.6 log cfu/g and 5.0 to 5.4 log cfu/g, respectively. The growth of R. oligosporus and the final pH (4.9) in barley tempeh were not significantly influenced by any of the LAB investigated.

Co-digestion of cultivated microalgae and sewage sludge from municipal waste water treatment.

In this study two wet microalgae cultures and one dried microalgae culture were co-digested in different proportions with sewage sludge in mesophilic and thermophilic conditions. The aim was to evaluate if the co-digestion could lead to an increased efficiency of methane production compared to digestion of sewage sludge alone. The results showed that co-digestion with both wet and dried microalgae, in certain proportions, increased the biochemical methane potential (BMP) compared with digestion of sewage sludge alone in mesophilic conditions. The BMP was significantly higher than the calculated BMP in many of the mixtures. This synergetic effect was statistically significant in a mixture containing 63% (w/w VS based) undigested sewage sludge and 37% (w/w VS based) wet algae slurry, which produced 23% more methane than observed with undigested sewage sludge alone. The trend was that thermophilic co-digestion of microalgae and undigested sewage sludge did not give the same synergy.

Impact of trace element addition on biogas production from food industrial waste--linking process to microbial communities.

Laboratory-scale reactors treating food industry waste were used to investigate the effects of additions of cobalt (Co), nickel/molybdenum/boron (Ni/Mo/B) and selenium/tungsten (Se/W) on the biogas process and the associated microbial community. The highest methane production (predicted value: 860 mL g(-1) VS) was linked to high Se/W concentrations in combination with a low level of Co. A combination of quantitative real-time PCR of 16S rRNA genes, terminal restriction fragment length polymorphism (T-RFLP) and clone library sequencing was used for the community analysis. The T-RFLP data show a higher diversity for bacteria than for archaea in all the treatments. The most abundant bacterial population (31-55% of the total T-RFLP fragments' intensity) was most closely related to Actinomyces europaeus (94% homology). Two dominant archaeal populations shared 98-99% sequence homology with Methanosarcina siciliae and Methanoculleus bourgensis, respectively. Only limited influence of the trace metal additions was found on the bacterial community composition, with two bacterial populations responding to the addition of a combination of Ni/Mo/B, while the dominant archaeal populations were influenced by the addition of Ni/Mo/B and/or Se/W. The maintenance of methanogenic activity was largely independent of archaeal community composition, suggesting a high degree of functional redundancy in the methanogens of the biogas reactors.

Rhizopus oligosporus and yeast co-cultivation during barley tempeh fermentation--nutritional impact and real-time PCR quantification of fungal growth dynamics.

Barley tempeh was produced by fermenting barley kernels with Rhizopus oligosporus. The potential of the yeasts Saccharomyces cerevisiae (three strains), S. boulardii (one strain), Pichia anomala (one strain) and Kluyveromyces lactis (one strain) to grow together with R. oligosporus during barley tempeh fermentation was evaluated. All yeast strains grew during the fermentation and even during cold storage of tempeh (P<0.01). The growth of yeasts slightly increased the ergosterol contents, but did not influence amino acid contents and compositions, and did not reduce phytate contents. Slight increases of vitamins B(6) and niacinamide, and slight decreases of B(1) and biotin were observed. Quantification of fungal growth is difficult during mixed species fermentations because ergosterol is found in all fungal species, and colony-forming-unit (cfu) estimations are not reliable for R. oligosporus and other sporulating fungi. Therefore, we developed a quantitative real-time PCR method for individually quantifying S. cerevisiae and R. oligosporus growth in barley tempeh. The PCR results were highly correlated with the ergosterol content of R. oligosporus and with the number of cfu of S. cerevisiae. Thus, real-time PCR is a rapid and selective method to quantify yeasts and R. oligosporus during mixed species fermentation of inhomogenous substrate such as barley tempeh.